masters term paper

How to Write a Term Paper From Start to Finish

The term paper, often regarded as the culmination of a semester's hard work, is a rite of passage for students in pursuit of higher education. Here's an interesting fact to kick things off: Did you know that the term paper's origins can be traced back to ancient Greece, where scholars like Plato and Aristotle utilized written works to explore and document their philosophical musings? Just as these great minds once wrote their thoughts on parchment, you, too, can embark on this intellectual voyage with confidence and skill.

How to Write a Term Paper: Short Description

In this article, we'll delve into the core purpose of this kind of assignment – to showcase your understanding of a subject, your research abilities, and your capacity to communicate complex ideas effectively. But it doesn't stop there. We'll also guide you in the art of creating a well-structured term paper format, a roadmap that will not only keep you on track but also ensure your ideas flow seamlessly and logically. Packed with valuable tips on writing, organization, and time management, this resource promises to equip you with the tools needed to excel in your academic writing.

Understanding What Is a Term Paper

A term paper, a crucial component of your college education, is often assigned towards the conclusion of a semester. It's a vehicle through which educators gauge your comprehension of the course content. Imagine it as a bridge between what you've learned in class and your ability to apply that knowledge to real-world topics.

For instance, in a history course, you might be asked to delve into the causes and consequences of a significant historical event, such as World War II. In a psychology class, your term paper might explore the effects of stress on mental health, or in an environmental science course, you could analyze the impact of climate change on a specific region.

Writing a term paper isn't just about summarizing facts. It requires a blend of organization, deep research, and the art of presenting your findings in a way that's both clear and analytical. This means structuring your arguments logically, citing relevant sources, and critically evaluating the information you've gathered.

For further guidance, we've prepared an insightful guide for you authored by our expert essay writer . It's brimming with practical tips and valuable insights to help you stand out in this academic endeavor and earn the recognition you deserve.

How to Start a Term Paper

Before you start, keep the guidelines for the term paper format firmly in mind. If you have any doubts, don't hesitate to reach out to your instructor for clarification before you begin your research and writing process. And remember, procrastination is your worst enemy in this endeavor. If you're aiming to produce an exceptional piece and secure a top grade, it's essential to plan ahead and allocate dedicated time each day to work on it. Now, let our term paper writing services provide you with some valuable tips to help you on your journey:

• Hone Your Topic : Start by cultivating a learning mindset that empowers you to effectively organize your thoughts. Discover how to research a topic in the section below.
• Hook Your Readers: Initiate a brainstorming session and unleash a barrage of creative ideas to captivate your audience right from the outset. Pose intriguing questions, share compelling anecdotes, offer persuasive statistics, and more.
• Craft a Concise Thesis Statement Example : If you find yourself struggling to encapsulate the main idea of your paper in just a sentence or two, it's time to revisit your initial topic and consider narrowing it down.
• Understand Style Requirements: Your work must adhere to specific formatting guidelines. Delve into details about the APA format and other pertinent regulations in the section provided.
• Delve Deeper with Research : Equipped with a clearer understanding of your objectives, dive into your subject matter with a discerning eye. Ensure that you draw from reputable and reliable sources.
• Begin Writing: Don't obsess over perfection from the get-go. Just start writing, and don't worry about initial imperfections. You can always revise or remove those early sentences later. The key is to initiate the term papers as soon as you've amassed sufficient information.

Ace your term paper with EssayPro 's expert help. Our academic professionals are here to guide you through every step, ensuring your term paper is well-researched, structured, and written to the highest standards.

Term Paper Topics

Selecting the right topic for your term paper is a critical step, one that can significantly impact your overall experience and the quality of your work. While instructors sometimes provide specific topics, there are instances when you have the freedom to choose your own. To guide you on how to write a term paper, consider the following factors when deciding on your dissertation topics :

• Relevance to Assignment Length: Begin by considering the required length of your paper. Whether it's a substantial 10-page paper or a more concise 5-page one, understanding the word count will help you determine the appropriate scope for your subject. This will inform whether your topic should be broad or more narrowly focused.
• Availability of Resources : Investigate the resources at your disposal. Check your school or community library for books and materials that can support your research. Additionally, explore online sources to ensure you have access to a variety of reference materials.
• Complexity and Clarity : Ensure you can effectively explain your chosen topic, regardless of how complex it may seem. If you encounter areas that are challenging to grasp fully, don't hesitate to seek guidance from experts or your professor. Clarity and understanding are key to producing a well-structured term paper.
• Avoiding Overused Concepts : Refrain from choosing overly trendy or overused topics. Mainstream subjects often fail to captivate the interest of your readers or instructors, as they can lead to repetitive content. Instead, opt for a unique angle or approach that adds depth to your paper.
• Manageability and Passion : While passion can drive your choice of topic, it's important to ensure that it is manageable within the given time frame and with the available resources. If necessary, consider scaling down a topic that remains intriguing and motivating to you, ensuring it aligns with your course objectives and personal interests.

Worrying About the Quality of Your Upcoming Essay?

"Being highly trained professionals, our writers can provide term paper help by creating a paper specifically tailored to your needs.

Term Paper Outline

Before embarking on the journey of writing a term paper, it's crucial to establish a well-structured outline. Be mindful of any specific formatting requirements your teacher may have in mind, as these will guide your outline's structure. Here's a basic format to help you get started:

• Cover Page: Begin with a cover page featuring your name, course number, teacher's name, and the deadline date, centered at the top.
• Abstract: Craft a concise summary of your work that informs readers about your paper's topic, its significance, and the key points you'll explore.
• Introduction: Commence your term paper introduction with a clear and compelling statement of your chosen topic. Explain why it's relevant and outline your approach to addressing it.
• Body: This section serves as the meat of academic papers, where you present the primary findings from your research. Provide detailed information about the topic to enhance the reader's understanding. Ensure you incorporate various viewpoints on the issue and conduct a thorough analysis of your research.
• Results: Share the insights and conclusions that your research has led you to. Discuss any shifts in your perspective or understanding that have occurred during the course of your project.
• Discussion: Conclude your term paper with a comprehensive summary of the topic and your findings. You can wrap up with a thought-provoking question or encourage readers to explore the subject further through their own research.

How to Write a Term Paper with 5 Steps

Before you begin your term paper, it's crucial to understand what a term paper proposal entails. This proposal serves as your way to introduce and justify your chosen topic to your instructor, and it must gain approval before you start writing the actual paper.

In your proposal, include recent studies or research related to your topic, along with proper references. Clearly explain the topic's relevance to your course, outline your objectives, and organize your ideas effectively. This helps your instructor grasp your term paper's direction. If needed, you can also seek assistance from our expert writers and buy term paper .

Draft the Abstract

The abstract is a critical element while writing a term paper, and it plays a crucial role in piquing the reader's interest. To create a captivating abstract, consider these key points from our dissertation writing service :

• Conciseness: Keep it short and to the point, around 150-250 words. No need for lengthy explanations.
• Highlight Key Elements: Summarize the problem you're addressing, your research methods, and primary findings or conclusions. For instance, if your paper discusses the impact of social media on mental health, mention your research methods and significant findings.
• Engagement: Make your abstract engaging. Use language that draws readers in. For example, if your paper explores the effects of artificial intelligence on the job market, you might begin with a question like, 'Is AI revolutionizing our work landscape, or should we prepare for the robots to take over?'
• Clarity: Avoid excessive jargon or technical terms to ensure accessibility to a wider audience.

Craft the Introduction

The introduction sets the stage for your entire term paper and should engage readers from the outset. To craft an intriguing introduction, consider these tips:

• Hook Your Audience: Start with a captivating hook, such as a thought-provoking question or a compelling statistic. For example, if your paper explores the impact of smartphone addiction, you could begin with, 'Can you remember the last time you went a whole day without checking your phone?'
• State Your Purpose: Clearly state the purpose of your paper and its relevance. If your term paper is about renewable energy's role in combating climate change, explain why this topic is essential in today's world.
• Provide a Roadmap: Briefly outline how your paper is structured. For instance, if your paper discusses the benefits of mindfulness meditation, mention that you will explore its effects on stress reduction, emotional well-being, and cognitive performance.
• Thesis Statement: Conclude your introduction with a concise thesis statement that encapsulates the central argument or message of your paper. In the case of a term paper on the impact of online education, your thesis might be: 'Online education is revolutionizing learning by providing accessibility, flexibility, and innovative teaching methods.'

Develop the Body Sections: Brainstorming Concepts and Content

Generate ideas and compose text: body sections.

The body of your term paper is where you present your research, arguments, and analysis. To generate ideas and write engaging text in the body sections, consider these strategies from our research paper writer :

• Structure Your Ideas: Organize your paper into sections or paragraphs, each addressing a specific aspect of your topic. For example, if your term paper explores the impact of social media on interpersonal relationships, you might have sections on communication patterns, privacy concerns, and emotional well-being.
• Support with Evidence: Back up your arguments with credible evidence, such as data, research findings, or expert opinions. For instance, when discussing the effects of social media on mental health, you can include statistics on social media usage and its correlation with anxiety or depression.
• Offer Diverse Perspectives: Acknowledge and explore various viewpoints on the topic. When writing about the pros and cons of genetic engineering, present both the potential benefits, like disease prevention, and the ethical concerns associated with altering human genetics.
• Use Engaging Examples: Incorporate real-life examples to illustrate your points. If your paper discusses the consequences of climate change, share specific instances of extreme weather events or environmental degradation to make the topic relatable.
• Ask Thought-Provoking Questions: Integrate questions throughout your text to engage readers and stimulate critical thinking. In a term paper on the future of artificial intelligence, you might ask, 'How will AI impact job markets and the concept of work in the coming years?'

Formulate the Conclusion

The conclusion section should provide a satisfying wrap-up of your arguments and insights. To craft a compelling term paper example conclusion, follow these steps:

• Revisit Your Thesis: Begin by restating your thesis statement. This reinforces the central message of your paper. For example, if your thesis is about the importance of biodiversity conservation, reiterate that biodiversity is crucial for ecological balance and human well-being.
• Summarize Key Points: Briefly recap the main points you've discussed in the body of your paper. For instance, if you've been exploring the impact of globalization on local economies, summarize the effects on industries, job markets, and cultural diversity.
• Emphasize Your Main Argument: Reaffirm the significance of your thesis and the overall message of your paper. Discuss why your findings are important or relevant in a broader context. If your term paper discusses the advantages of renewable energy, underscore its potential to combat climate change and reduce our reliance on fossil fuels.
• Offer a Thoughtful Reflection: Share your own reflections or insights about the topic. How has your understanding evolved during your research? Have you uncovered any unexpected findings or implications? If your paper discusses the future of space exploration, consider what it means for humanity's quest to explore the cosmos.
• End with Impact: Conclude your term paper with a powerful closing statement. You can leave the reader with a thought-provoking question, a call to action, or a reflection on the broader implications of your topic. For instance, if your paper is about the ethics of artificial intelligence, you could finish by asking, 'As AI continues to advance, what ethical considerations will guide our choices and decisions?'

Edit and Enhance the Initial Draft

After completing your initial draft, the revision and polishing phase is essential for improving your paper. Here's how to refine your work efficiently:

• Take a Break: Step back and return to your paper with a fresh perspective.
• Structure Check: Ensure your paper flows logically and transitions smoothly from the introduction to the conclusion.
• Clarity and Conciseness: Trim excess words for clarity and precision.
• Grammar and Style: Proofread for errors and ensure consistent style.
• Citations and References: Double-check your citations and reference list.
• Peer Review: Seek feedback from peers or professors for valuable insights.
• Enhance Intro and Conclusion: Make your introduction and conclusion engaging and impactful.
• Coherence Check: Ensure your arguments support your thesis consistently.
• Read Aloud: Reading your paper aloud helps identify issues.
• Final Proofread: Perform a thorough proofread to catch any remaining errors.

Term Paper Format

When formatting your term paper, consider its length and the required citation style, which depends on your research topic. Proper referencing is crucial to avoid plagiarism in academic writing. Common citation styles include APA and MLA.

If unsure how to cite term paper for social sciences, use the APA format, including the author's name, book title, publication year, publisher, and location when citing a book.

For liberal arts and humanities, MLA is common, requiring the publication name, date, and location for referencing.

Adhering to the appropriate term paper format and citation style ensures an organized and academically sound paper. Follow your instructor's guidelines for a polished and successful paper.

Term Paper Example

To access our term paper example, simply click the button below.

The timeline of events from 1776 to 1861, that, in the end, prompted the American Civil War, describes and relates to a number of subjects modern historians acknowledge as the origins and causes of the Civil War. In fact, pre-Civil War events had both long-term and short-term influences on the War—such as the election of Abraham Lincoln as the American president in 1860 that led to the Fall of Fort Sumter in April of the same year. In that period, contentions that surrounded states’ rights progressively exploded in Congress—since they were the initial events that formed after independence. Congress focused on resolving significant issues that affected the states, which led to further issues. In that order, the US’s history from 1776 to 1861 provides a rich history, as politicians brought forth dissimilarities, dissections, and tensions between the Southern US & the people of slave states, and the Northern states that were loyal to the Union. The events that unfolded from the period of 1776 to 1861 involved a series of issues because they promoted the great sectional crisis that led to political divisions and the build-up to the Civil War that made the North and the South seem like distinctive and timeless regions that predated the crisis itself.

Final Thoughts

In closing, approach the task of writing term papers with determination and a positive outlook. Begin well in advance, maintain organization, and have faith in your capabilities. Don't hesitate to seek assistance if required, and express your individual perspective with confidence. You're more than capable of succeeding in this endeavor!

Need a Winning Hand in Academia?

Arm yourself with our custom-crafted academic papers that are sharper than a well-honed pencil! Order now and conquer your academic challenges with style!

What is the Difference between a Term Paper and a Research Paper?

What is the fastest way to write a term paper, related articles.

• PRO Courses Guides New Tech Help Pro Expert Videos About wikiHow Pro Upgrade Sign In
• EDIT Edit this Article
• EXPLORE Tech Help Pro About Us Random Article Quizzes Request a New Article Community Dashboard This Or That Game Popular Categories Arts and Entertainment Artwork Books Movies Computers and Electronics Computers Phone Skills Technology Hacks Health Men's Health Mental Health Women's Health Relationships Dating Love Relationship Issues Hobbies and Crafts Crafts Drawing Games Education & Communication Communication Skills Personal Development Studying Personal Care and Style Fashion Hair Care Personal Hygiene Youth Personal Care School Stuff Dating All Categories Arts and Entertainment Finance and Business Home and Garden Relationship Quizzes Cars & Other Vehicles Food and Entertaining Personal Care and Style Sports and Fitness Computers and Electronics Health Pets and Animals Travel Education & Communication Hobbies and Crafts Philosophy and Religion Work World Family Life Holidays and Traditions Relationships Youth
• Browse Articles
• Learn Something New
• Quizzes Hot
• This Or That Game New
• Train Your Brain
• Explore More
• Support wikiHow
• About wikiHow
• Log in / Sign up
• Education and Communications
• College University and Postgraduate
• Academic Writing
• Research Papers

Everything You Need to Know to Write an A+ Term Paper

Last Updated: March 4, 2024 Fact Checked

Sample Term Papers

Researching & outlining.

• Drafting Your Paper
• Revising Your Paper

Expert Q&A

This article was co-authored by Matthew Snipp, PhD and by wikiHow staff writer, Raven Minyard, BA . C. Matthew Snipp is the Burnet C. and Mildred Finley Wohlford Professor of Humanities and Sciences in the Department of Sociology at Stanford University. He is also the Director for the Institute for Research in the Social Science’s Secure Data Center. He has been a Research Fellow at the U.S. Bureau of the Census and a Fellow at the Center for Advanced Study in the Behavioral Sciences. He has published 3 books and over 70 articles and book chapters on demography, economic development, poverty and unemployment. He is also currently serving on the National Institute of Child Health and Development’s Population Science Subcommittee. He holds a Ph.D. in Sociology from the University of Wisconsin—Madison. There are 13 references cited in this article, which can be found at the bottom of the page. This article has been fact-checked, ensuring the accuracy of any cited facts and confirming the authority of its sources. This article has been viewed 2,228,746 times.

A term paper is a written assignment given to students at the end of a course to gauge their understanding of the material. Term papers typically count for a good percentage of your overall grade, so of course, you’ll want to write the best paper possible. Luckily, we’ve got you covered. In this article, we’ll teach you everything you need to know to write an A+ term paper, from researching and outlining to drafting and revising.

Quick Steps to Write a Term Paper

• Hook your readers with an interesting and informative intro paragraph. State your thesis and your main points.
• Support your thesis by providing quotes and evidence that back your claim in your body paragraphs.
• Summarize your main points and leave your readers with a thought-provoking question in your conclusion.

• Think of your term paper as the bridge between what you’ve learned in class and how you apply that knowledge to real-world topics.
• For example, a history term paper may require you to explore the consequences of a significant historical event, like the Civil War. An environmental science class, on the other hand, may have you examine the effects of climate change on a certain region.
• Your guidelines should tell you the paper’s word count and formatting style, like whether to use in-text citations or footnotes and whether to use single- or double-spacing. If these things aren’t specified, be sure to reach out to your instructor.

• Make sure your topic isn’t too broad. For example, if you want to write about Shakespeare’s work, first narrow it down to a specific play, like Macbeth , then choose something even more specific like Lady Macbeth’s role in the plot.
• If the topic is already chosen for you, explore unique angles that can set your content and information apart from the more obvious approaches many others will probably take. [3] X Research source
• Try not to have a specific outcome in mind, as this will close you off to new ideas and avenues of thinking. Rather than trying to mold your research to fit your desired outcome, allow the outcome to reflect a genuine analysis of the discoveries you made. Ask yourself questions throughout the process and be open to having your beliefs challenged.
• Reading other people's comments, opinions, and entries on a topic can often help you to refine your own, especially where they comment that "further research" is required or where they posit challenging questions but leave them unanswered.

• For example, if you’re writing a term paper about Macbeth , your primary source would be the play itself. Then, look for other research papers and analyses written by academics and scholars to understand how they interpret the text.

• For example, if you’re writing a paper about Lady Macbeth, your thesis could be something like “Shakespeare’s characterization of Lady Macbeth reveals how desire for power can control someone’s life.”
• Remember, your research and thesis development doesn’t stop here. As you continue working through both the research and writing, you may want to make changes that align with the ideas forming in your mind and the discoveries you continue to unearth.
• On the other hand, don’t keep looking for new ideas and angles for fear of feeling confined. At some point, you’re going to have to say enough is enough and make your point. You may have other opportunities to explore these questions in future studies, but for now, remember your term paper has a finite word length and an approaching due date!

• Abstract: An abstract is a concise summary of your paper that informs readers of your topic, its significance, and the key points you’ll explore. It must stand on its own and make sense without referencing outside sources or your actual paper.
• Introduction: The introduction establishes the main idea of your paper and directly states the thesis. Begin your introduction with an attention-grabbing sentence to intrigue your readers, and provide any necessary background information to establish your paper’s purpose and direction.
• Body paragraphs: Each body paragraph focuses on a different argument supporting your thesis. List specific evidence from your sources to back up your arguments. Provide detailed information about your topic to enhance your readers’ understanding. In your outline, write down the main ideas for each body paragraph and any outstanding questions or points you’re not yet sure about.
• Results: Depending on the type of term paper you’re writing, your results may be incorporated into your body paragraphs or conclusion. These are the insights that your research led you to. Here you can discuss how your perspective and understanding of your topic shifted throughout your writing process.
• Conclusion: Your conclusion summarizes your argument and findings. You may restate your thesis and major points as you wrap up your paper.

Drafting Your Term Paper

• Writing an introduction can be challenging, but don’t get too caught up on it. As you write the rest of your paper, your arguments might change and develop, so you’ll likely need to rewrite your intro at the end, anyway. Writing your intro is simply a means of getting started and you can always revise it later. [10] X Trustworthy Source PubMed Central Journal archive from the U.S. National Institutes of Health Go to source
• Be sure to define any words your readers might not understand. For example, words like “globalization” have many different meanings depending on context, and it’s important to state which ones you’ll be using as part of your introductory paragraph.

• Try to relate the subject of the essay (say, Plato’s Symposium ) to a tangentially related issue you happen to know something about (say, the growing trend of free-wheeling hookups in frat parties). Slowly bring the paragraph around to your actual subject and make a few generalizations about why this aspect of the book/subject is so fascinating and worthy of study (such as how different the expectations for physical intimacy were then compared to now).

• You can also reflect on your own experience of researching and writing your term paper. Discuss how your understanding of your topic evolved and any unexpected findings you came across.

• While peppering quotes throughout your text is a good way to help make your point, don’t overdo it. If you use too many quotes, you’re basically allowing other authors to make the point and write the paper for you. When you do use a quote, be sure to explain why it is relevant in your own words.
• Try to sort out your bibliography at the beginning of your writing process to avoid having a last-minute scramble. When you have all the information beforehand (like the source’s title, author, publication date, etc.), it’s easier to plug them into the correct format.

Revising & Finalizing Your Term Paper

• Trade in weak “to-be” verbs for stronger “action” verbs. For example: “I was writing my term paper” becomes “I wrote my term paper.”

• It’s extremely important to proofread your term paper. If your writing is full of mistakes, your instructor will assume you didn’t put much effort into your paper. If you have too many errors, your message will be lost in the confusion of trying to understand what you’ve written.

• If you add or change information to make things clearer for your readers, it’s a good idea to look over your paper one more time to catch any new typos that may have come up in the process.

• The best essays are like grass court tennis—the argument should flow in a "rally" style, building persuasively to the conclusion. Thanks Helpful 0 Not Helpful 0
• If you get stuck, consider giving your professor a visit. Whether you're still struggling for a thesis or you want to go over your conclusion, most instructors are delighted to help and they'll remember your initiative when grading time rolls around. Thanks Helpful 0 Not Helpful 0
• At least 2 hours for 3-5 pages.
• At least 4 hours for 8-10 pages.
• At least 6 hours for 12-15 pages.
• Double those hours if you haven't done any homework and you haven't attended class.
• For papers that are primarily research-based, add about two hours to those times (although you'll need to know how to research quickly and effectively, beyond the purview of this brief guide).

You Might Also Like

• ↑ https://www.binghamton.edu/counseling/self-help/term-paper.html
• ↑ Matthew Snipp, PhD. Research Fellow, U.S. Bureau of the Census. Expert Interview. 26 March 2020.
• ↑ https://emory.libanswers.com/faq/44525
• ↑ https://writing.wisc.edu/handbook/assignments/planresearchpaper/
• ↑ https://owl.purdue.edu/owl/general_writing/the_writing_process/thesis_statement_tips.html
• ↑ https://libguides.usc.edu/writingguide/outline
• ↑ https://gallaudet.edu/student-success/tutorial-center/english-center/writing/guide-to-writing-introductions-and-conclusions/
• ↑ https://www.ncbi.nlm.nih.gov/pubmed/26731827
• ↑ https://writing.wisc.edu/handbook/assignments/writing-an-abstract-for-your-research-paper/
• ↑ https://www.ivcc.edu/stylesite/Essay_Title.pdf
• ↑ https://www.uni-flensburg.de/fileadmin/content/institute/anglistik/dokumente/downloads/how-to-write-a-term-paper-daewes.pdf
• ↑ https://library.sacredheart.edu/c.php?g=29803&p=185937
• ↑ https://www.cornerstone.edu/blog-post/six-steps-to-really-edit-your-paper/

About This Article

If you need to write a term paper, choose your topic, then start researching that topic. Use your research to craft a thesis statement which states the main idea of your paper, then organize all of your facts into an outline that supports your thesis. Once you start writing, state your thesis in the first paragraph, then use the body of the paper to present the points that support your argument. End the paper with a strong conclusion that restates your thesis. For tips on improving your term paper through active voice, read on! Did this summary help you? Yes No

• Send fan mail to authors

Reader Success Stories

Bill McReynolds

Apr 7, 2017

Did this article help you?

Gerard Mortera

Mar 30, 2016

Ayuba Muhammad Bello

Dec 28, 2016

Mar 24, 2016

Jera Andarino

May 11, 2016

Featured Articles

Trending Articles

Watch Articles

• Terms of Use
• Privacy Policy
• Do Not Sell or Share My Info
• Not Selling Info

Don’t miss out! Sign up for

wikiHow’s newsletter

• The Difference Between a Master’s Thesis and a Term Paper

Navigating the world of graduate studies can be a challenging adventure, filled with numerous academic milestones. One question that often pops up is: what's the difference between a Master's thesis and a term paper? While both are crucial elements in your educational journey, understanding their distinct roles, structures, and purposes is key to successfully fulfilling their requirements. Whether you're seeking Master's thesis help or simply want to understand how a term paper differs from a Master thesis, this blog post will serve as your guide. We'll explore the nuances of each, giving you a clearer picture of what to expect and how to excel in both.

Level of Education

When it comes to the level of education, a Master's thesis and a term paper serve different educational tiers and signify different milestones in your academic journey. A term paper is commonly associated with undergraduate studies or early graduate coursework . It's usually a requirement for a specific class and aims to demonstrate your understanding of the subject matter covered throughout the semester. The emphasis is often on synthesizing existing knowledge rather than creating new insights. You're expected to research a topic, usually of your choosing within the scope of the course, and present your findings in a well-organized manner. However, term papers usually don't require the depth of research or the lengthy process that a Master's thesis demands.

On the other hand, a Master's thesis is a staple of graduate education, specifically aimed at Master's level students . If you're on the journey toward obtaining your Master's degree, crafting a thesis will likely be one of your final academic obligations before graduation. This is a long-form research project that requires a profound understanding of your chosen field. Unlike a term paper, a Master's thesis expects you to contribute new knowledge or perspectives to your area of study. Given the scope and depth, many students seek Master's thesis help to guide them through the complex process of research, writing, and defense.

In summary, while term papers are often shorter, topic-focused papers you'll encounter throughout your educational journey, a Master's thesis is a much more comprehensive project, usually reserved for the conclusion of your Master's program. Knowing these distinctions can greatly help you in setting the right expectations and seeking the appropriate guidance, whether that's Master thesis help for the graduate students among us, or simply understanding the requirements for a solid term paper.

Scope and Depth

Another significant difference between a Master's thesis and a term paper lies in their scope and depth. Simply put, the Master's thesis is a marathon, while the term paper is more like a sprint.

A term paper is generally confined to the subject matter of a specific course or semester. You pick a topic that interests you (within the boundaries of the course), do some research, and write a paper that ranges from about 10 to 20 pages , depending on the guidelines. The purpose is to show that you've understood the material and can apply your knowledge in a cohesive manner. It's relatively short-term, often taking a few weeks to complete.

In contrast, a Master's thesis is a far more involved process requiring a more extensive commitment to research and analysis. Typically, it's a document of 60 to 100 pages or more . Because it requires such a deep dive into a specialized topic, many students opt for Master's thesis help, from identifying a suitable research question to the final process of writing and defending the thesis. The goal of a Master's thesis is not just to show that you understand existing research but to contribute new insights to your field. It demands rigorous methodology, extensive literature reviews, and months, if not years, of focused work.

The Master's thesis allows, or rather requires, you to immerse yourself in a topic, often leading to publishable quality work. It often serves as a stepping stone to further research or a future Ph.D. This makes seeking Master thesis help particularly worthwhile for those looking to produce high-quality academic work that stands up to scrutiny.

So, when contemplating the scope and depth of your next academic project, remember that a term paper is often about learning and synthesizing existing knowledge quickly, while a Master's thesis is about making a meaningful, long-term contribution to your field. The kind of guidance or assistance you'll need—be it quick pointers or more substantial Master's thesis help—will depend on which type of project you're tackling.

Research Emphasis

The level of research required for a term paper versus a Master's thesis is another point of distinction that should not be overlooked. Both assignments involve research, of course, but the depth, methodology, and end goals can be vastly different.

For a term paper, you'll likely be drawing from existing academic articles, books, or other primary sources related to your topic. The focus here is often on understanding, explaining, and integrating existing theories or viewpoints. The ability to construct an argument or provide a new angle on a well-known topic is valued, but you aren't necessarily expected to contribute groundbreaking new information to your field. The research aspect here is substantial but usually not as intensive, making it unlikely that you'd need to seek out specialized term paper assistance.

In contrast, a Master's thesis places a strong emphasis on research that contributes something new to your academic field. This usually involves a detailed literature review, followed by original research—either qualitative or quantitative. You might conduct surveys, engage in interviews, or carry out experiments. Given the complex nature of this kind of research, many students look for Master's thesis help to navigate through designing experiments, understanding ethical considerations, or analyzing data.

It's not just about gathering information but about asking meaningful questions and developing theories or models that could prompt further research in your field. Because of these high expectations and complexities, it's not uncommon for students to seek Master thesis help, whether that's in the form of faculty mentorship, peer review, or specialized research assistance services.

So, in a nutshell, if you're writing a term paper, you're usually focused on synthesizing existing knowledge into a coherent narrative. If you're tackling a Master's thesis, you're diving deep into uncharted waters to bring something new to the surface. Your research is not just an academic exercise but a contribution to your field, making Master's thesis help not just helpful, but often essential.

Length and Format

The physical characteristics of a term paper and a Master's thesis, including their length and format, can also tell you a lot about the depth and complexity of each.

Term papers are often shorter, running between 10 to 20 pages depending on the course requirements. The format is usually simpler, sticking to academic norms such as introduction, body, and conclusion, along with standard citation styles like APA, MLA, or Chicago . While the term paper is often a solo endeavor, the goal is to demonstrate mastery of course material, and the focus is more on clarity, organization, and correct citation of sources.

Conversely, a Master's thesis is a much more substantial document. Length can vary by field and institution, but a typical thesis is often between 60 to 100 pages or even more. The format is far more formalized and is often divided into various sections like abstract, acknowledgment, table of contents, introduction, literature review, methodology, findings, discussion, conclusion, references, and appendices. Each of these sections has its own purpose and criteria for what should be included, adding layers of complexity that often prompt students to seek Master thesis help.

Given the comprehensive nature of a Master's thesis, the formatting and organization are crucial, not just for the presentation but also for the ease of future researchers who may read your work. Software for citation management, specialized statistical packages for data analysis , and even project management tools can be incredibly useful. All these complexities often make students search for Master's thesis help to ensure that their work meets academic standards and is free from errors in formatting and citation.

In summary, while term papers require you to present well-researched arguments in a straightforward way, Master's theses demand a more rigid structure and far greater length to discuss, analyze, and conclude your original research. Understanding the differences in the required length and format between a term paper and a Master's thesis is essential, and Master's thesis help can often be invaluable in navigating these requirements successfully.

Contribution to the Field

One of the most defining differences between a term paper and a Master's thesis is the expected contribution to the academic field. While both projects require a certain level of expertise, the weight they carry in the academic world varies greatly.

A term paper aims to demonstrate a student's understanding and interpretation of existing knowledge. While insightful perspectives and well-structured arguments in a term paper can be applauded, they are not designed to make a lasting contribution to the field. Term papers are seldom published and, while they can be incredibly valuable for your personal and academic development, their impact is typically confined to your course grade.

In stark contrast, a Master's thesis aims to contribute new knowledge, insights, or methodologies to an existing academic conversation . This doesn't mean that every Master's thesis revolutionizes its field, but the expectation is that the work should be of a quality that could be published in an academic journal. A well-executed Master's thesis can become a stepping-stone to a Ph.D. program, academic publications, or advanced-level employment within the field. Given these high stakes, many students opt for Master thesis help to ensure their work is of the highest quality and worthy of contributing to academic discourse.

Whether you are extending previous research, filling a gap in existing literature, or perhaps challenging established theories, your Master's thesis holds the potential to influence future research and even policy. Because of this, there are often more rigorous checks for quality, originality, and significance, making Master's thesis help an important consideration for ensuring your work stands up to such scrutiny.

To summarize, while term papers help to build your foundation in academic research and writing, a Master's thesis aims to add a building block to the existing structure of scholarly work in your field. The level of originality and scholarly contribution required for a Master's thesis is considerably higher, which is why professional Master's thesis help can be so invaluable for students aiming to make a lasting impact.

Evaluation and Assessment

The criteria for evaluating a term paper and a Master's thesis differ substantially, primarily due to their divergent purposes and scopes. Both are academic exercises, but they are assessed through lenses that consider their unique contributions to your educational journey.

Term papers are usually evaluated by the course instructor, and the assessment focuses on your understanding of the course material, quality of research, coherence of argument, and clarity of expression. In most cases, the term paper's grade will be one component of your overall course grade. Evaluation is typically quick, and you might receive feedback within weeks or even days. If you're struggling with the demands of a term paper, there are numerous resources for assistance, but given its limited scope, many students don't feel the need for specialized term paper help.

On the other hand, a Master's thesis undergoes a more stringent and extended evaluation process. It's not just about impressing one instructor but satisfying a committee of experts in the field. The thesis defense is a defining feature of this evaluative process. During this oral examination, you'll be required to answer questions and defend your research and conclusions.

The aim is not merely to pass but to excel, as the evaluation will influence not only your grade but your academic and possibly professional reputation. Because of the comprehensive and challenging nature of this evaluation process, many students seek Master thesis help to prepare for the rigorous scrutiny they will face.

Additionally, the Master's thesis often undergoes several rounds of revisions, based on feedback from advisors or the review committee, before it is accepted. Even after acceptance, it may be subject to peer review if submitted to an academic journal. The evaluation, therefore, doesn't end at graduation but can continue as the work is exposed to broader academic scrutiny. This ongoing process of evaluation is another reason why Master's thesis help can be so crucial, guiding students through not just the initial submission but any subsequent revisions or submissions to academic journals.

In conclusion, the assessment of a term paper tends to be quicker, less formal, and confined to the context of a specific course. In contrast, a Master's thesis goes through a rigorous, extended evaluation process with wider academic implications, often making Master's thesis help a worthwhile investment for those aiming for the highest standards of excellence.

Master's Thesis Help by BridgeText

Embarking on the journey of writing a Master's thesis presents numerous challenges, from selecting a topic and designing research to data collection, statistical analysis, and the writing process. Given the complexity and high stakes, it's unsurprising that many students seek specialized support. That's where BridgeText comes in, a trusted name in the academic writing sector with nearly three decades of experience. You can order thesis paper online now!

What sets us apart at BridgeText is our dedicated focus on thesis work. With 28 years in the field, we've fine-tuned our approach to cover every aspect of thesis writing, offering a holistic package for students. Our in-house statistics team is particularly beneficial for those tackling data-driven research. Unlike many other services that outsource this crucial element, we ensure that statistical integrity is maintained, thereby offering both quality and continuity in the research process.

Another reason to consider Master thesis help from BridgeText is our seasoned team of academic writers, all specialized in various disciplines. With this wealth of expertise, you're not just getting assistance; you're gaining a partner who understands the intricacies of your specific field. Our guidance extends beyond mere writing help, offering valuable inputs that can significantly elevate the quality of your work.

So, whether you're stuck on your research methodology or caught in the endless loop of revisions, we offer a suite of Master's thesis help services designed to navigate you through these challenges. Our aim is not just to help you pass but to excel, ensuring that your Master's thesis becomes a meaningful contribution to your field. In essence, BridgeText is more than a service provider; we're your academic ally, empowering you to produce a Master's thesis that withstands rigorous scrutiny and leaves a lasting impact.

By investing in specialized Master's thesis help like that offered by us at BridgeText, you're not merely purchasing a service; you're investing in peace of mind. With our in-depth expertise and long-standing experience, we provide an invaluable resource for any student aiming to submit a high-quality Master's thesis.

Recent Posts

Renaming Variables in Stata

Calculating the Binomial Confidence Interval in Stata

The Detection of AI-Generated Academic Text

Sorting Variables in Stata

Have any questions?

Our support team is ready to answer your questions.

Help Center FAQ  

To mark our first year, we've slashed all our prices in half. Order now to seize this limited opportunity!

                Place Your Order

How To Write a Term Paper: A Guide That Works

30 June, 2020

16 minutes read

Author:  Mathieu Johnson

Once you’ve started your university career, you are going to be asked to present a term paper. What’s the difference between a term paper and a research paper? How can you write a good term? What’s the best way to structure it? Where can you find some tips to make the writing process faster? In this article, we’ll discuss a few tips to help you prepare a term paper quickly and professionally.

What Is a Term Paper… And What Is The First Step?

A term paper is a critical and analytical report on the topic or subject that you covered within the course of studies. It usually consists of two separate but equally important aspects: your own thoughts about the topic and a demonstration of your understanding of the existing literature. The main goal of this assignment is to summarize the material you learned and showcase your understanding of the topic. This aspect makes the term paper a universal instrument for assessing a student’s proficiency. It also explains why term papers cost so many points of your course grade.

We usually associate a term paper with a research paper , but although the concepts are quite similar, a research paper requires a more academic approach and a deeper investigation into the literature of your field of study.

To write an outstanding college term paper, you must understand that your professor has requested it in order to test your analytical thinking skills. You must collect relevant data, analyze it, and then make a summary or solve a particular problem. Such skills are highly relevant to the business world, so this type of the task is as practical as it is educational.

So, let’s start the preparation!

Before you begin writing

Unfortunately, there is no magical recipe that allows you to get everything done fast. You will need to choose the best way forward in whatever situation you find yourself, but here are some tips to help you prepare for the assignment.

To begin with, take the research stage seriously . Sometimes, when students are really interested in a topic, they only want to present their personal ideas about the problem. Unfortunately, if you’re not completely familiar with all the data from the various sources, you will need to reinvent the bicycle.

Term paper writing was never an easy ride. Well, not for our expert writers. Place an order with our term paper writing service and secure yourself an “A!”

In the initial stages of your research, investigate everything you can find on the topic . This may sound like a tall order, but you’ll find that it doesn’t actually entail that much reading. At this point you are only compiling the research, so you will be skimming through numerous prospects rather than reading them completely. Bear in mind that your aim is to get acquainted with the various aspects of your problem. The term paper summarizes the knowledge you gained within a course and requires to familiarize yourself with the research that other people have already made on your topic.

Thinking that your opinions are completely original and unique is quite egocentric, and it can get you into trouble. So, “your” thoughts about the problem are usually just somebody else’s statements that you have rephrased (or even a well-established academic concept!). Remember that your professor will be familiar with all the literature surrounding the issue: if you merely rewrite someone else’s thoughts and present them as your own (even if you don’t realize doing it), be prepared for criticism!

Applying a Structure To Your Term Paper

Once you have read all the leading authors and their approaches to your problem, it’s time to create a structure for your work. This is not yet an outline; you just need to decide what to write about. Sketch out the topic for the theoretical portion of your work and think about practical aspects and how you can approach the research in the best possible way.

At this point, you really need to call or email your supervisor . Your professor will have seen hundreds of term papers like yours (i.e., they have not yet been written, but a definite idea exists!) and will be prepared to give you feedback and advice. He or she will tell you what literature you have omitted, offer suggestions about what you should read, and give you feedback about your paper. It may well be that your approach has already occurred to somebody else, in which case there is no need to repeat it.

Choosing a Topic: Easy as Riding a Bike?

When you choose your topic, make sure you choose something that you are interested in . That’s our advice if you want a painless term paper. If you prefer to investigate a field that you’ve never really explored before, you can challenge yourself to do that, too. That might be sophisticated, but why not?

If you decide to investigate a topic or a problem that you are pretty familiar with, your writing will be more fluid. You will focus your attention on a specific aspect of the chosen field and expand your knowledge within that scope. On the contrary, choosing an unfamiliar subject matter can wash out your expertise.

Be prepared to change the topic if you find out that your research isn’t going anywhere. It might occur that you presuppose that your topic has a potential but somewhere at the stage of initial research, you find that it just won’t work. It’s always a good idea to consider two or three topics when you kick off the term paper writing – even if they are just different ways of examining the same problem. By doing this, you will be able to choose the best version, which may not be the one you started with at all!

Related Post: 100 Persuasive essay topics

Formulating a Thesis statement

Writing a proper thesis statement can also be challenging. To begin with, write down a couple of prominent ideas or concepts, then try to make rough drafts of them to see how they’ll work in the structural framework. You will probably find that one idea fits your style, interests, and knowledge base: you can choose that one as your thesis statement.

Remember that the thesis statement is the skeleton, the central concept of your paper. It is the elemental attribute of almost any academic paper – from master’s thesis to a simple five paragraph essay. If you do a thorough job on it, you will find that writing (and defending!) your argument is much easier.

Be aware that all of these stages are parts of a procedure – one leads to another. When writing a term paper, you should collect the material and wrap it up at the same time.

Planning – The Key To Success

Some people claim that they can write a term paper without any planning. In our opinion, this is impossible. If you don’t have a postgraduate degree and you aren’t a certified genius, you need to prepare an outline for your project. It may come as a surprise, but even people who claim otherwise actually prepare outlines – in their heads. But if you don’t have that much experience, use a pencil and your notebook to ensure that you don’t forget anything.

Don’t procrastinate on your College or University papers anymore. Get professional help with our essay writer !

That’s when we get to preparing your first draft . There’s only one thing to add here: do as many drafts as you need in order to achieve your goal. Understand that your aim is to create an excellent term paper and keep working at it until you are satisfied.

Term Paper Outline: Write Everything In The Proper Section!

In the Introduction , state the topic that you are going to investigate and the context of your work. This is the critical ‘selling’ moment of your work. In a nutshell, your introduction combined with a conclusion should give a sneak peek into what the whole paper is about. If your introduction is well-prepared, it will be quite complacent about the body of your project. The introduction must include an abstract that presents your thesis statement . You should explain your motivation (why should the reader be concerned about this problem?) , your methods (what scientific tools did you use?) , and the results (what you achieved) .

The Literature Review totally corresponds to its name – it is here to review the literature you compiled. Your professor will double check it to make sure that you understand the context of your argument. One more thing to add is: collect all the information you can! Ideally, you should read or at least glance through every book and author that you can find on the topic. Think of your task as a fascinating journey: if you approach it like that, reading hundreds of pages won’t seem like that much of a challenge.

In the Discussion , you must present the interpretations of the problem. Be honest, explain what you pieces of data you don’t agree with and what ideas and concepts you support. This section connects the dots between theory and practice when writing a term paper. Wherever possible, provide several interpretations of the subject matter, then choose the one(s) that are most relevant to the case you are presenting.

In the Body , focus on those arguments that prove your thesis statement. This section must be absolutely logical. If you have chosen a more complicated topic, use heading and sub-headings to improve the appearance of this section. While writing the body, keep your target audience (your professors) in mind. In other words, don’t just record the obvious causes/effects/solutions but also showcase your own findings – what you have discovered and how that proves your thesis statement. Demonstrate that you are familiar with the details and you will stun your readers with the prolific mastery of the topic.

Now, the Conclusion   is her to summarize both the content and the purpose of the paper. The most challenging part is not to make it too dry. Reiterate your thesis statement and briefly show how your results justified your proposition. At the very end, you can suggest a call to action or pose a rhetorical question or statement that leaves your reader wanting more.

What to do next?

When you have finished, reread your work a couple of times. You will almost certainly find a few faults, whether they are contextual, factual, syntactical, grammatical, or even simple spelling mistakes. A very useful tip is to wait for two or three days after writing your final draft to proofread it afterward. Your brain will have time to process the information, and you’ll be able to look at it with a fresh view.

When proofreading, take care to polish the structural problems. The skeleton (the logic and the thesis statement) should make sense. If they don’t, try to approach the problem from another perspective. The changes may take some time, but bear in mind that your objective is to produce professional work. Be patient!

After that, print the term paper. The human eye processes information differently on the paper than on a computer screen; that’s why you need to print it and take one final look for any possible mistakes. Even if you don’t see any serious defects, pay attention to formatting, punctuation, and synonyms. It’s an academic text, so make it shine!

Term Paper Sample

Be sure to check the sample of a term paper, completed by our writers. Use it as an example to perfect your own writing. Link:  Term Paper Sample: Consumer Buying Behavior .

The Do’s and Don’ts of Term Paper Writing

There you have the most important tips to help you succeed in writing a term paper. Now it’s up to you to stop reading and start writing!

A life lesson in Romeo and Juliet taught by death

Due to human nature, we draw conclusions only when life gives us a lesson since the experience of others is not so effective and powerful. Therefore, when analyzing and sorting out common problems we face, we may trace a parallel with well-known book characters or real historical figures. Moreover, we often compare our situations with […]

Ethical Research Paper Topics

Writing a research paper on ethics is not an easy task, especially if you do not possess excellent writing skills and do not like to contemplate controversial questions. But an ethics course is obligatory in all higher education institutions, and students have to look for a way out and be creative. When you find an […]

Art Research Paper Topics

Students obtaining degrees in fine art and art & design programs most commonly need to write a paper on art topics. However, this subject is becoming more popular in educational institutions for expanding students’ horizons. Thus, both groups of receivers of education: those who are into arts and those who only get acquainted with art […]

How to Write a Term Paper in 5 Steps

Term papers are a key way to test a student’s knowledge and research skills, but they can be difficult to write. In this guide, we explain the best methods to write a term paper, including the proper term paper format and even how to choose a term paper topic.

Cite your sources with confidence Grammarly helps you avoid plagiarism Write with Grammarly

What is a term paper?

A term paper is a piece of academic writing in which a student demonstrates their knowledge of a topic of study. Term papers constitute a large portion of the final grade, making them a serious assignment. There is typically no more than one term paper assigned each term, although how long a term lasts depends on the school system.

Keep in mind that a term paper is one specific type of academic paper. It is more intensive than a standard writing assignment but is not as in-depth as a thesis paper or dissertation.

How long is a term paper?

There is no standard length for a term paper; each subject, course, and professor has their own preferences. Term papers can be as short as five pages or as long as twenty pages, but they usually fall somewhere in the middle.

What’s the difference between a term paper and a research paper?

Technically speaking, a research paper is a paper that argues its main point with original data and evidence. However, the term research paper is used informally to refer to any paper that requires research, even when collecting data and evidence from other preexisting sources. So in that sense, a term paper can be a research paper if the student must research other sources to complete it.

The terms term paper and research paper are often used interchangeably. However, term papers are generally assigned once per term, whereas a teacher or professor can assign as many research papers as they wish.

What’s the difference between a term paper and an essay?

An essay is any writing that asserts the author’s opinion or perspective, whether for school, publication, or just the author’s personal enjoyment. Unlike research-oriented term papers that draw from data and evidence, essay writing is based only on the author’s experience or viewpoint.

Essays are usually shorter than term papers and more casual in tone. Keep in mind that term papers are strictly academic, whereas essays can be written for various audiences.

How do I write a term paper?

Writing a term paper still follows the standard writing process but with some extra focus in certain areas.

1 Developing ideas

The first step of writing a term paper is brainstorming to come up with potential topics and then selecting the best one. Sometimes your topics are assigned, but often you’ll have to choose one yourself.

In addition to picking a topic that you’re personally interested in, try to settle on one that has sufficient depth. Avoid topics that are too broad because you won’t be able to cover everything, and stay away from topics that are too specific because you may not find enough information to fill the required paper length.

If you’re looking for inspiration, check out our list of term and research paper topics .

2 Preparation (research)

The preparation stage is when you determine your main point and the parts of your topic you’re going to discuss. For most term papers, that requires research. If you’re not conducting your own research, then you’re finding and reviewing sources to use instead.

A good place to start is by writing your thesis statement , a single sentence that sums up the main point(s) your paper tries to make. Your thesis statement determines what evidence and counterarguments you’ll need to discuss. Deciding on these early can help streamline your research.

Once you establish what you want to include in your term paper, you can start putting it in order by writing an outline . Think of the outline as the blueprint of your term paper, mapping out each part of your topic, paragraph by paragraph.

Be sure to follow the term paper format for the assignment. This means adhering to the guidelines and planning enough content to meet the length requirement.

4 First draft

Writing the first draft is easier if you follow your outline. Although this stage can be the most labor-intensive, remember that everything doesn’t need to be perfect. You can still go back later to revise and optimize your wording, but for the first draft, just focus on getting all your ideas down on paper.

This isn’t always easy. If you’re having trouble or get stuck at certain points, go back to the fundamentals and revisit your first-year writing skills. If you have writer’s block, don’t be afraid to take a break and try again later—your brain could just be too tired to come up with ideas.

5 Editing and proofreading

After you have completed a first draft, it’s time to begin the editing process. This is when you correct the mistakes in the first draft and detect other issues that need revising. If a section seems weak or inadequate, you can revise the wording or even rewrite it entirely. You may find that something is missing from your first draft, so now is the time to add it.

We recommend rereading your term paper twice—once to correct the wording and structural mistakes and another time to proofread . Revising it twice allows you to better focus on particular issues instead of trying to address everything at once. If you’re trying to determine the right word choice , spending time on spelling and grammar might be a distraction. It’s better to separate the tasks and do them one at a time.

Term paper FAQs

How do i write my term paper.

Writing a term paper still follows the standard writing process, but goes deeper into certain areas. Start by brainstorming topics that you find interesting before selecting one that has ample source material. Then begin your research. When you’re ready to start writing, create an outline, then a first draft, and finally revisions.

There is no standard length for a term paper; every teacher or professor has their own requirements. Term papers can be as short as five pages or as long as twenty pages, but they usually fall somewhere in the middle.

Technically speaking, a research paper supports its thesis with original data and evidence. However, the term research paper is used informally to refer to any paper that requires research, even when collecting data and evidence from other preexisting sources. So in that sense, a term paper can also be a research paper if the student relies on other sources to complete it.

Find Study Materials for

• Business Studies
• Combined Science
• Computer Science
• Engineering
• English Literature
• Environmental Science
• Human Geography
• Macroeconomics
• Microeconomics
• Social Studies
• Browse all subjects
• Read our Magazine

Create Study Materials

Term paper – master your fear of writing.

Just be honest and tell me, how many words? And how much time have you got to do it? If you’re about to embark on writing a term paper, I have excellent news for you – this is a crash course! I’m about to introduce you to the fine art of writing term papers for any topic. We will look at an airtight outline example, some life hacks, and the usual formatting tips. Let’s dive right in!

What Is a Term Paper? Definition and Guidelines

If you’ve only just started university and have already been slammed by this frightful word, welcome to this new level of study! University is not only about cramming from tons of books: It should also, ideally, foster critical thinking, teach you how to argue your points effectively, and help you develop research skills. And you will need all three of these to write a stellar term paper!

But hang on a sec, what is a term paper? A term paper is a longer type of research-based homework on a particular topic. Term papers range from 15 to 25 pages because any less is considered lazy and any more is too much for any professor to read (trust me, I teach at a university).

In general, you should be free to select a topic for your term paper, but regardless of whether you’re free to do it or are assigned one, term papers mostly have the same goal. Namely, they test your ability to formulate and support your arguments and locate yourself in a particular theoretical framework. Sound scary yet? Don’t worry! I’m here to illuminate some of the vaguer aspects of term paper writing.

Join over 90% of students getting better grades!

That’s a pretty good statistic. Download our free all-in-one learning app and start your most successful learning journey yet. Let’s do it!

Structuring Your Term Paper Outline (+ Sample Term Paper Outline PDF)

Before you begin writing, it’s advisable to have an idea about where exactly your writing is going. The best way to achieve this is to write an outline, or (as we sleep-deprived academics like to call it) an abstract. An abstract is a short description of your paper/article/project that outlines your main research questions and the theoretical framework you will be working with.

I generally suggest that people start with a very simple pyramid structure when writing an abstract:

• The foundation. This is where you introduce a broad, general statement on the topic of your choice. You can clarify and specify this in a few more sentences to ease your readers into the research project.

Example: Contemporary drama boasts the power to transform the audience through careful selection and crafty delivery of impactful images. By creating faux-reality, drama sometimes appeals to the affective side of the audience in order to provide commentary on a number of social and psychological issues. Duncan Macmillan’s Every Brilliant Thing capitalises on its affect-inducing potential, tackling the issue of suicidal depression.

• The middle. In this part of the outline, you state the aims of your study. Some of my favourite phrases to signal your intentions include: this paper aims to shed light on, the goal of this research paper is, the idea behind this term paper is, etc. Feel free to add some powerful verbs of action such as examine, assess, illuminate, discern, analyse, cross-reference, etc. to emphasise your ideas.

Example: This paper aims to explore how the play creates a more realistic setting by deviating from the audience’s expectations, thus blurring the line between drama and real life. It may be argued that simulated reality, exemplified through a number of exaggerations, impacts the affective component in the audience’s attitude formation and that its neglect of the cognitive reinforces the transformative power of Every Brilliant Thing .

• The top. The final part of your outline should highlight coherent hypotheses or research questions that your study will answer. While academic papers usually dream of some originality, this should not concern you yet – you don’t need to invent hot water in your term papers, but as you gain experience, novel conclusions will become easier to form.

Example: This paper will then take a final look at how the structure of the play simulates depression in order to sensitise the audience and to which extent it attains its goal of conveying the message of the universality and repercussions of the disease.

Writing an outline is a good way to organise your thoughts, figure out what kind of books you need, and anticipate your results.

In the abovementioned examples, the books you’d need would have to do with theatre, psychological influences, and simulation of reality.

This process applies to any subject. The outline can be more detailed, but it needn’t go over 300 words. A word of advice: if you cannot summarise the key points of your topic in 300 words, you should do some more brainstorming until you reach the specific goal.

PS Check out this excellent term paper outline sample !

We offer more, for less. Way less.

Our all-in-one learning platform is completely free. Access teacher-verified explanations, flashcards with spaced repetition, and a study planner with analytics. At no cost to you.

Term Paper Format – The Safety Net

Each term paper should follow a relatively familiar structure and way of arguing your points. Let’s start with the basics:

• Cover page. This is where your title goes (centred, bolded, pt24 ideally). The cover page should also list your personal details, such as name, address, email, student ID number, phone number (maybe), and the institution and the department for which you are writing your term paper. Each university tends to have its own layout for the cover page, but the rule of thumb is that institutional information goes above the title, whereas personal information is below.
• Table of contents – your readers need to know what to expect!
• Introduction. This is a more elaborate version of your project outline. You should specify what the paper is dealing with, what theoretical framework you’re using, and what your hypothesis is. My pro tip is to write the introduction last because term papers tend to grow as you write and you may end up with vastly different results from those you had expected.
• Theoretical framing. Explain which theories or ideas you’re using.
• Methodology. This is mostly present in scientific papers where you must explain what methods will guide your study (i.e. experiments).
• Analysis. Close readings, experiments, data surveys – whatever your project is doing, it should be doing it here.
• Discussion . Feel free to start interpreting your results in this section. A great paper does not simply list data – it compares and contrasts. You must be able to draw conclusions about what your analysis has shown you. Results as expected? Hypothesis confirmed. Results not ideal? There’s something to write about. Consider why something turned out differently and what that means for future studies.
• Optional: pitfalls and future improvements. Again, this is more present in sciences than humanities, but you could address possible pitfalls or blind spots in your study and suggest how they can be improved upon in the future. You can also talk about what lines of research your project can inspire.
• Conclusion . Time to wrap it all up. Briefly summarise the key points of research and main results. If you haven’t already devoted a separate section of the paper to this, you can also write about indications for future research in your conclusion.

Term Paper Structure Example

To give you a more precise example of a structured term paper, here’s a more detailed structure of the above-described example on theatre:

Still Unsure about How to Write a Term Paper?

Excellent, I love good questions! The truth is, writing a term paper is a labour of love (it is hard labour, especially if you’re carrying all the books!), so I will give you some tips on how to make it an enjoyable experience.

• Pick a topic you’re interested in. There’s nothing you can say to convince me that your subject is so absolutely wretchedly uninteresting that you simply cannot find such a topic. You just haven’t done your work yet. Start digging and follow the internet clicking abyss until you stumble upon something that takes your fancy. My master thesis idea was based on a single line I read in a magazine about Neil Gaiman’s American Gods – I managed to turn it into 80 pages, two scientific articles, and two talks just fine, even though it may not have been researched previously. So, whatever you’re writing about, there’s got to be a fun angle to it.
• Start reading. You cannot write a term paper from nothing. Once you have a general topic and an outline, you should start collecting your materials. Check out your library and inform yourself about the inter-library loan. Get acquainted with various scientific databases like JSTOR and ResearchGate – your university probably has wide access to many knowledge repositories you can use through an official VPN or library computers. Search by keywords and titles and save everything that sounds interesting. Learn to recognise important elements and ideas in those texts and be ready to use them to support your arguments.
• Know when to stop, too. Sometimes you’ll find yourself deep in the excitement of learning something new, but there will come a point when you realise you’re ready to put what you’ve found into your own words. Set up an experiment, survey, or study and follow up on the results. In humanities, this may mean a closer analysis of selected texts. This is where you start writing – again, leave the introduction for later and jump right into the core of the work.
• Mind the style. When writing a term paper, you need to keep certain standards up. Term papers are written in the ‘academic’ style and involve lots of passive voice, verbs of enlightenment (illustrate, examine, assess), and words marking cause-effect relationships. Don’t be afraid to use transition words to make your text and conclusions flow easily.
• Cite properly! Oh, how I hated learning all the citation styles when I was just starting out, but once you do learn the ropes, it gets easier. It’s a bit of drudgery, but my advice is to write down your sources meticulously as you go along. As soon as you cite someone, make sure you add the full citation at the end of the text (I like having them in a separate document), and don’t forget in-text citations. Depending on what field you’re studying in, you will have different citation styles (like MLA, Chicago, APA) at your disposal – make sure you check the requirements for each course and consult the corresponding websites with guidelines.

Change the way you learn, starting right now!

With thousands of teacher-reviewed explanations and flashcards on all your subjects, you have everything you need to succeed!

Don’t Forget about Term Paper Editing!

And there goes the last-minute churning of text and hitting send before passing out for the next two days. Writing a term paper at university should not be left for the last minute. If you’re a chronic procrastinator, it’s time to learn to organise your time and devote enough of it to your assignments.

When you’re done with writing, you should leave your paper alone for a few days – sleep on it, as they say. You can treat this distance like any good study break – it’ll help you clear your mind, prevent resentment towards the subject, and allow you to see it through new eyes. Before submitting, re-read your text carefully and edit the writing. Weed out spelling and grammatical errors and prune unnecessary examples or repetitive statements. A good way to do this is to change the font or even font size in your writing software – this engages your perception and makes spotting mistakes easier.

Editing is also the time to consider how your arguments are holding together and whether you need to add or replace some text and/or rearrange your points. It’s an extremely important part of the writing process, but you shouldn’t overdo it either. Perfectionism can get you into the editing spiral that usually leads to messing up parts that were initially good. A few re-reads are fine, but anything more and you might as well start to rewrite the whole thing.

The last question to consider is whether you are happy with your result. Remember, this is a term paper and you’re still learning, so nobody expects it to be perfect, but you should be satisfied with what you’ve accomplished.

The Key Takeaways of Writing a Term Paper

Writing a term paper is a longer commitment than a simple essay. To ensure your success, start well ahead of time or you might find yourself rushed and stressed .

• Try to find a topic of personal interest to you.
• Scribble an outline to work out your angle or general direction of the paper.
• Read enough materials. Your library and online databases are your friends.
• Form hypotheses and set up experiments or analyses.
• Get down to business (and stop procrastinating !).
• Don’t forget to edit the paper well and format it correctly.

Source: Danica Stojanovic, ‘Theatrical (Hyper)Reality: The Effects of Breaking Formal Boundaries in Every Brilliant Thing ’, Over The Horizon, London, 2020, pp. 81‑100.

Did you know that StudySmarter was rated best study app worldwide!

What is a term paper?

How do you write a term paper, how do you write a term paper proposal, how do you write a term paper outline, how long should a term paper be, how do you prepare a term paper, how do you write a conclusion for a term paper, how do you write a term paper introduction.

Have a language expert improve your writing

Run a free plagiarism check in 10 minutes, generate accurate citations for free.

• Knowledge Base
• Dissertation
• What Is a Thesis? | Ultimate Guide & Examples

What Is a Thesis? | Ultimate Guide & Examples

Published on September 14, 2022 by Tegan George . Revised on November 21, 2023.

A thesis is a type of research paper based on your original research. It is usually submitted as the final step of a master’s program or a capstone to a bachelor’s degree.

Writing a thesis can be a daunting experience. Other than a dissertation , it is one of the longest pieces of writing students typically complete. It relies on your ability to conduct research from start to finish: choosing a relevant topic , crafting a proposal , designing your research , collecting data , developing a robust analysis, drawing strong conclusions , and writing concisely .

Thesis template

You can also download our full thesis template in the format of your choice below. Our template includes a ready-made table of contents , as well as guidance for what each chapter should include. It’s easy to make it your own, and can help you get started.

Download Word template Download Google Docs template

Instantly correct all language mistakes in your text

Upload your document to correct all your mistakes in minutes

Table of contents

Thesis vs. thesis statement, how to structure a thesis, acknowledgements or preface, list of figures and tables, list of abbreviations, introduction, literature review, methodology, reference list, proofreading and editing, defending your thesis, other interesting articles, frequently asked questions about theses.

You may have heard the word thesis as a standalone term or as a component of academic writing called a thesis statement . Keep in mind that these are two very different things.

• A thesis statement is a very common component of an essay, particularly in the humanities. It usually comprises 1 or 2 sentences in the introduction of your essay , and should clearly and concisely summarize the central points of your academic essay .
• A thesis is a long-form piece of academic writing, often taking more than a full semester to complete. It is generally a degree requirement for Master’s programs, and is also sometimes required to complete a bachelor’s degree in liberal arts colleges.
• In the US, a dissertation is generally written as a final step toward obtaining a PhD.
• In other countries (particularly the UK), a dissertation is generally written at the bachelor’s or master’s level.

The only proofreading tool specialized in correcting academic writing - try for free!

The academic proofreading tool has been trained on 1000s of academic texts and by native English editors. Making it the most accurate and reliable proofreading tool for students.

Try for free

The final structure of your thesis depends on a variety of components, such as:

• Your discipline
• Your theoretical approach

Humanities theses are often structured more like a longer-form essay . Just like in an essay, you build an argument to support a central thesis.

In both hard and social sciences, theses typically include an introduction , literature review , methodology section ,  results section , discussion section , and conclusion section . These are each presented in their own dedicated section or chapter. In some cases, you might want to add an appendix .

Thesis examples

We’ve compiled a short list of thesis examples to help you get started.

• Example thesis #1:   “Abolition, Africans, and Abstraction: the Influence of the ‘Noble Savage’ on British and French Antislavery Thought, 1787-1807” by Suchait Kahlon.
• Example thesis #2: “’A Starving Man Helping Another Starving Man’: UNRRA, India, and the Genesis of Global Relief, 1943-1947″ by Julian Saint Reiman.

The very first page of your thesis contains all necessary identifying information, including:

• Your full title
• Your full name
• Your department
• Your institution and degree program
• Your submission date.

Sometimes the title page also includes your student ID, the name of your supervisor, or the university’s logo. Check out your university’s guidelines if you’re not sure.

Read more about title pages

The acknowledgements section is usually optional. Its main point is to allow you to thank everyone who helped you in your thesis journey, such as supervisors, friends, or family. You can also choose to write a preface , but it’s typically one or the other, not both.

Read more about acknowledgements Read more about prefaces

Receive feedback on language, structure, and formatting

Professional editors proofread and edit your paper by focusing on:

• Academic style
• Vague sentences
• Style consistency

See an example

An abstract is a short summary of your thesis. Usually a maximum of 300 words long, it’s should include brief descriptions of your research objectives , methods, results, and conclusions. Though it may seem short, it introduces your work to your audience, serving as a first impression of your thesis.

Read more about abstracts

A table of contents lists all of your sections, plus their corresponding page numbers and subheadings if you have them. This helps your reader seamlessly navigate your document.

Your table of contents should include all the major parts of your thesis. In particular, don’t forget the the appendices. If you used heading styles, it’s easy to generate an automatic table Microsoft Word.

Read more about tables of contents

While not mandatory, if you used a lot of tables and/or figures, it’s nice to include a list of them to help guide your reader. It’s also easy to generate one of these in Word: just use the “Insert Caption” feature.

Read more about lists of figures and tables

If you have used a lot of industry- or field-specific abbreviations in your thesis, you should include them in an alphabetized list of abbreviations . This way, your readers can easily look up any meanings they aren’t familiar with.

Read more about lists of abbreviations

Relatedly, if you find yourself using a lot of very specialized or field-specific terms that may not be familiar to your reader, consider including a glossary . Alphabetize the terms you want to include with a brief definition.

Read more about glossaries

An introduction sets up the topic, purpose, and relevance of your thesis, as well as expectations for your reader. This should:

• Ground your research topic , sharing any background information your reader may need
• Define the scope of your work
• Introduce any existing research on your topic, situating your work within a broader problem or debate
• State your research question(s)
• Outline (briefly) how the remainder of your work will proceed

In other words, your introduction should clearly and concisely show your reader the “what, why, and how” of your research.

Read more about introductions

A literature review helps you gain a robust understanding of any extant academic work on your topic, encompassing:

• Selecting relevant sources
• Determining the credibility of your sources
• Critically evaluating each of your sources
• Drawing connections between sources, including any themes, patterns, conflicts, or gaps

A literature review is not merely a summary of existing work. Rather, your literature review should ultimately lead to a clear justification for your own research, perhaps via:

• Addressing a gap in the literature
• Building on existing knowledge to draw new conclusions
• Exploring a new theoretical or methodological approach
• Introducing a new solution to an unresolved problem
• Definitively advocating for one side of a theoretical debate

Read more about literature reviews

Theoretical framework

Your literature review can often form the basis for your theoretical framework, but these are not the same thing. A theoretical framework defines and analyzes the concepts and theories that your research hinges on.

Read more about theoretical frameworks

Your methodology chapter shows your reader how you conducted your research. It should be written clearly and methodically, easily allowing your reader to critically assess the credibility of your argument. Furthermore, your methods section should convince your reader that your method was the best way to answer your research question.

A methodology section should generally include:

• Your overall approach ( quantitative vs. qualitative )
• Your research methods (e.g., a longitudinal study )
• Your data collection methods (e.g., interviews or a controlled experiment
• Any tools or materials you used (e.g., computer software)
• The data analysis methods you chose (e.g., statistical analysis , discourse analysis )
• A strong, but not defensive justification of your methods

Read more about methodology sections

Your results section should highlight what your methodology discovered. These two sections work in tandem, but shouldn’t repeat each other. While your results section can include hypotheses or themes, don’t include any speculation or new arguments here.

Your results section should:

• State each (relevant) result with any (relevant) descriptive statistics (e.g., mean , standard deviation ) and inferential statistics (e.g., test statistics , p values )
• Explain how each result relates to the research question
• Determine whether the hypothesis was supported

Additional data (like raw numbers or interview transcripts ) can be included as an appendix . You can include tables and figures, but only if they help the reader better understand your results.

Read more about results sections

Your discussion section is where you can interpret your results in detail. Did they meet your expectations? How well do they fit within the framework that you built? You can refer back to any relevant source material to situate your results within your field, but leave most of that analysis in your literature review.

For any unexpected results, offer explanations or alternative interpretations of your data.

Read more about discussion sections

Your thesis conclusion should concisely answer your main research question. It should leave your reader with an ultra-clear understanding of your central argument, and emphasize what your research specifically has contributed to your field.

Why does your research matter? What recommendations for future research do you have? Lastly, wrap up your work with any concluding remarks.

Read more about conclusions

In order to avoid plagiarism , don’t forget to include a full reference list at the end of your thesis, citing the sources that you used. Choose one citation style and follow it consistently throughout your thesis, taking note of the formatting requirements of each style.

Which style you choose is often set by your department or your field, but common styles include MLA , Chicago , and APA.

Create APA citations Create MLA citations

In order to stay clear and concise, your thesis should include the most essential information needed to answer your research question. However, chances are you have many contributing documents, like interview transcripts or survey questions . These can be added as appendices , to save space in the main body.

Read more about appendices

Once you’re done writing, the next part of your editing process begins. Leave plenty of time for proofreading and editing prior to submission. Nothing looks worse than grammar mistakes or sloppy spelling errors!

Consider using a professional thesis editing service or grammar checker to make sure your final project is perfect.

Once you’ve submitted your final product, it’s common practice to have a thesis defense, an oral component of your finished work. This is scheduled by your advisor or committee, and usually entails a presentation and Q&A session.

After your defense , your committee will meet to determine if you deserve any departmental honors or accolades. However, keep in mind that defenses are usually just a formality. If there are any serious issues with your work, these should be resolved with your advisor way before a defense.

If you want to know more about AI for academic writing, AI tools, or research bias, make sure to check out some of our other articles with explanations and examples or go directly to our tools!

Research bias

• Survivorship bias
• Self-serving bias
• Availability heuristic
• Halo effect
• Hindsight bias
• Deep learning
• Generative AI
• Machine learning
• Reinforcement learning
• Supervised vs. unsupervised learning

 (AI) Tools

• Grammar Checker
• Paraphrasing Tool
• Text Summarizer
• AI Detector
• Plagiarism Checker
• Citation Generator

The conclusion of your thesis or dissertation shouldn’t take up more than 5–7% of your overall word count.

If you only used a few abbreviations in your thesis or dissertation , you don’t necessarily need to include a list of abbreviations .

If your abbreviations are numerous, or if you think they won’t be known to your audience, it’s never a bad idea to add one. They can also improve readability, minimizing confusion about abbreviations unfamiliar to your reader.

When you mention different chapters within your text, it’s considered best to use Roman numerals for most citation styles. However, the most important thing here is to remain consistent whenever using numbers in your dissertation .

A thesis or dissertation outline is one of the most critical first steps in your writing process. It helps you to lay out and organize your ideas and can provide you with a roadmap for deciding what kind of research you’d like to undertake.

Generally, an outline contains information on the different sections included in your thesis or dissertation , such as:

• Your anticipated title
• Your abstract
• Your chapters (sometimes subdivided into further topics like literature review , research methods , avenues for future research, etc.)

A thesis is typically written by students finishing up a bachelor’s or Master’s degree. Some educational institutions, particularly in the liberal arts, have mandatory theses, but they are often not mandatory to graduate from bachelor’s degrees. It is more common for a thesis to be a graduation requirement from a Master’s degree.

Even if not mandatory, you may want to consider writing a thesis if you:

• Plan to attend graduate school soon
• Have a particular topic you’d like to study more in-depth
• Are considering a career in research
• Would like a capstone experience to tie up your academic experience

Cite this Scribbr article

If you want to cite this source, you can copy and paste the citation or click the “Cite this Scribbr article” button to automatically add the citation to our free Citation Generator.

George, T. (2023, November 21). What Is a Thesis? | Ultimate Guide & Examples. Scribbr. Retrieved April 9, 2024, from https://www.scribbr.com/dissertation/thesis/

Is this article helpful?

Tegan George

Other students also liked, dissertation & thesis outline | example & free templates, writing strong research questions | criteria & examples, 10 research question examples to guide your research project, what is your plagiarism score.

• Apply Apply (link opens in new window)
• Request Info
• Applied Nutrition
• Healthcare Administration
• Health Informatics
• Public Health
• Social Work
• Science Prerequisites
• UNE's Awards & Recognition
• Application Tips
• Resources for Students
• Faculty Development
• Resources, Links & News
• Alumni Spotlights
• Ambassador Spotlights
• Faculty Spotlights
• Student Spotlights
• Team Spotlights

How to Write Excellent Graduate-Level Papers

“How to Write Excellent Graduate-Level Papers” brought to you by the Student Academic Success Center (SASC) at UNE.

Becoming a better writer – the process

Breaking a writing project down into phases helps with motivation as well as managing your time and workload effectively. The phases of the process – prewriting, drafting, revision, and editing – are described below. Each step allows you to focus your energy in a particular way, with it all adding up to a more thoughtful, clear piece of writing.

The phases don’t have to be done in a set, linear order, if that’s not effective for you. If you like to write some rough draft paragraphs first, then go back and do a post-draft outline or revise those paragraphs before continuing, that’s fine. The key is to make sure each part of the process is done thoroughly before you consider your paper finished.

Let’s start with using prewriting to get the process rolling:

Using various prewriting strategies can help you avoid procrastinating and start a draft on the right track. You aren’t under pressure to develop a paper yet – this is about unlocking the flow of ideas. Play around with some of these strategies to find ones that work best for you:

• Tap into your curiosity

When you’re faced with an assignment, spend some time simply wondering about the topic. What intrigues you? Why should you and others in your profession care about it? Come up with a couple of relevant questions that you want to explore. Then consider which questions are most meaningful to you personally and professionally—and why? This can be done on paper, in conversation with someone else, or internally.

• Relate the assignment to your profession

Think about why the assignment is important to your field of study and work as a health professional, a social worker, an educator, etc. Making your assignment as personally and professionally relevant as possible helps with generating the motivation to start writing and keeping the momentum through the process. View this as an opportunity to learn useful information.

• Use the assignment itself as an outline

Copy the assignment and paste it into a new document. Break it apart visually by adding line spaces and/or tabs. This will help you more easily identify key concepts which need to be explained and verbs that indicate critical thinking is required (e.g., analyze, compare, evaluate). Create a rough outline using parts of the assignment as headings for different sections of the paper.

Similarly, you could annotate the assignment by marking up the key words and concepts and making little notes in the margins about what to add or how sections or ideas might tie together.

• Leverage what you already know, and then research with a purpose

Another very helpful strategy is to identify key concepts in the assignment description, then brainstorm what you already know about them based on the class readings or videos. Next, make a list of questions you still have about the concepts and overall topic. These will help drive the additional research needed to fill in your gaps of knowledge and locate credible evidence to support your explanations.

Having those questions makes researching more efficient because you have a purpose for reading: you’re looking for pieces of information rather than simply reading articles.

Read more: Faculty Spotlight: Lori Rand, Writing Specialist at SASC

The drafting phase involves determining your focus and starting to develop paragraph ideas within a structure. Keep a copy of the assignment on your draft as you write. Clarify the point of your paper – what is the main question that the assignment asking you to answer?

Think of a draft as packaging ideas into paragraphs that all relate to the paper’s main focus, as summed up in the thesis statement. For clarity, try to keep each paragraph focused on one idea at a time. However, because this phase is about getting thoughts down, and thoughts often jump around, drafting tends to be messy. That’s okay! The next step, revision, is where you really improve the writing.

In this phase, you can work on improving how you are guiding your reader through your thinking. Your reader will understand your ideas more easily if they are clearly focused, well-developed with specific evidence (correctly cited), and nicely organized.

Two strategies to guide you through revision include SASC’s Revision Checklist and Post-draft Outline, found here under Writing Resources. A writing appointment is also a great way to learn about and practice revision skills.

Editing is the final, polishing phase; it involves correcting sentence-level issues and technical aspects, such as word choice and grammar. Readers pick up these issues quickly because they can be the most obvious. Carelessness with grammar or word choice can lead to misunderstandings and make your writing seem unprofessional.

Trust the process

As mentioned earlier, the writing process is not necessarily a linear, step-by-step approach; it’s recursive, so it’s highly likely you’ll move back and forth between phases as you figure out your focus and organization of ideas.

Using this process gets easier with practice, and it works well in any writing situations, not just for graduate school assignments and scholarly papers.

Once you develop the most efficient method for your learning style, not only will you get faster, you will produce better academic papers.

Book an appointment

The SASC can help with all phases of the writing process via an Online Writing Support Appointment.  Visit the Online Student page for more details about writing support and resources.

For more online education insider tips and guides, subscribe today!

Weekly insights, best practices, student spotlights & more, straight to your inbox.

How to Write a Term Paper With Examples and Tips

11 December 2023

last updated

Students in higher learning institutions must submit their term papers at the end of each semester. Basically, these papers play a crucial role in evaluating the learner’s knowledge of a specific subject. In this case, scholars should engage in adequate preparation before writing a complete term paper. Then, some of the essential steps include defining a topic, finding credible sources, creating and revising a paper’s outline, and drafting a term work. Moreover, an outline of a term paper differs from that of other essays since it must include subsections. Further on, writers should ensure that all the subtopic relates to a thesis statement. Besides, each body paragraph must contain a topic sentence, supportive proof, appropriate descriptions, and a concluding and transitioning statement. In turn, the term paper’s conclusion should include a concise summary of the main points discussed in such work. Hence, students need to learn how to write a term paper to pass their academic goals.

Definition of a Term Paper

Students must prepare research papers for them to succeed in their studies. For example, a term paper refers to a serious research paper that a student should submit at the end of a semester. In this case, professors use these works to track and evaluate their learners’ knowledge about the area of expertise. Moreover, the process of organizing a term paper involves comprehensive research and methodological writing skills. Then, outstanding term papers contain analytical and organized structures. Besides, they have well-researched evidence that supports significant claims. In turn, learners in higher educational institutions prepare term papers when reflecting on their knowledge in a specific study area.

Possible Topics for Term Papers

Students in higher learning institutions may come across different topics for writing their term papers. In practice, term paper themes vary from one subject to another and require one to engage in detailed research. Hence, possible topics for writing a term paper that one may come across are:

• Is there a cancer epidemic due to industrial chemicals in the environment?
• Should federal courts be bound by the “original intent” of the framers?
• Do foreign investments threaten U.S. economic independence?
• Should morality and human rights influence foreign trade policy?
• Do rich nations have a responsibility to help developing countries?
• Partnership benefits at state and federal institutions.
• Same-sex adoption and access to reproductive technologies.
• Execution of juveniles.
• The lengthy appealing process for death row inmates.
• The Constitutional question of “cruel and unusual punishment.”

Term paper topics have a broad scope. Basically, term paper themes given above show that a writer must carry out extensive research to provide a comprehensive response. Also, students decide on the content that they must include in their term papers to give a comprehensive analysis of a subject.

Step-by-Step Guide for Writing a Term Paper

A student must prepare a term paper to achieve desired grades and complete a study course. Basically, adequate preparation allows scholars to gather relevant evidence and draft a term paper effectively. Hence, the necessary steps in writing a term paper that one should take to organize an academic piece are:

Step 1: Preparation

A. defining a topic for a term paper.

Defining a specific subject for a term paper is the first and most crucial activity that a writer must consider. For instance, describing an issue allows scholars to understand their course prompts and understand key ideas required to complete a term paper. In this case, one must understand the meaning of essential terms with the paper’s context. Moreover, students should use resources, like a dictionary and thesaurus, to obtain the necessary definitions. In turn, writers may opt to seek help from peers and lecturers when defining a topic for a term paper.   

B. Preparing Ideas

Preparing ideas for a term paper leads to outstanding works. For example, writers should identify all the relevant ideas and points needed to be covered before engaging in the actual writing process. Unfortunately, many learners fail to consider preparing thoughts as an essential step when writing a term paper. As a result, they prepare low-quality papers and achieve low grades. In turn, students need to redo their papers to pass their classes. Hence, writers should prepare ideas for writing a term paper by using acceptable methods.

C. Brainstorming

Scholars should consider brainstorming as an acceptable method to prepare ideas for a term paper. For example, brainstorming helps learners to come up with fresh and new facts. In this case, students think of the ideas that relate to their research topics. Moreover, the process generates unique ideas that can make one’s work to stand out. Hence, some ideas for brainstorming that one may use when preparing concepts and thoughts for term papers are:

• Come up with bad ideas first – Learners should think about research concepts related to their topics. In this case, successful students do not consider raising positive thoughts during brainstorming. Instead, they should present both good and bad ideas concerning their subjects. Also, writers should not feel stupid for raising bad ideas since the strategy helps identify weaker and more robust ideas. Then, one should allow ideas to flow during the brainstorming strategy. Besides, scholars should focus on raising positive opinions after exhausting throwaway thoughts. Hence, authors should increase both bad and good ideas that relate to their research topics.
• Breaking and building ideas – One of the most effective strategies of turning a few ideas into many is to break them down. Basically, learners should identify general themes that relate to a term paper and break them into smaller details. In this case, the process helps authors to see if some narrower ideas branch from main themes. Alternatively, one may combine different ideas to create a broader subtopic for a term paper. Hence, writers should break down more general concepts while combining narrow ones.
• Play word games – Outstanding term papers contain original and unique ideas. For example, word games are instrumental tools that prevent learners from producing generic and unoriginal ideas. In this case, word games motivate some out-of-box thinking. Moreover, “word storm” is an excellent method for a student to generate related ideas. In turn, this method allows authors to create thoughts naturally without overthinking.
• Creating a mood board – Learners should rely on methods that motivate them to generate fresh and unique ideas related to a research topic. For instance, combining imagery, color, and visual-spatial elements evoke emotions and feelings that spark fresh and new thoughts. In this case, students manage to recall some concepts acquired during learning by improving the term paper’s quality.
• Doodling – Successful students spur creativity insights and increase attention when generating essential ideas for a term paper. Basically, doodling allows a learner to engage with visuals that spark new thoughts. Also, practical doodling approaches help authors to break out of the traditional brainstorming approaches that rely on reading and talking. In this case, learners should break visual objects into small objects or combine unrelated items. Hence, these approaches motivate the brain to generate unique ideas for supporting a central theme.
• Changing a physical environment – Ordinary motivation plays a crucial role in the generation of new ideas. For example, students should change the physical environment to avoid boredom. In this case, enriched and attractive environments affect how the human brain works and speeds up how one generates new ideas and thoughts. In turn, a successful learner must select the location for brainstorming effectively. 

Reading is an appropriate method that students may consider when generating ideas for term papers. For instance, reading is a traditional method that writers use to raise arguments related to a specific topic. In this case, scholars should identify credible sources that relate to a research topic and read them to understand an assigned subject better. Also, this strategy plays a crucial in raising viable and accurate ideas about the term paper’s topic. However, scholars must take the necessary precautions since extensive reading is a tedious and monotonous process.   

E. Considering an Academic Audience

Different scholars read term papers for specific reasons. Basically, students must consider the target audience as academic readers to ensure that term papers meet their needs. In most cases, writers must use the official language when expressing thoughts. Moreover, formal language suits academic documents because it reveals professionalism and academic excellence.

Step 2: Setting Up the Stage

A. researching for sources.

Terms papers must contain credible evidence obtained from academic sources . Basically, scholars must gather adequate evidence from different reliable sources , like books, journal articles, financial and laboratory reports, credible websites, and magazines. As a ground rule, all sources must provide adequate and irrefutable evidence to support the main arguments. Also, one should find scholarly sources published in the last ten years because they contain the latest evidence and facts on research issues. Hence, writers should look for credible sources to support the main arguments in their term papers.

B. Making Notes for a Term Paper

Taking notes is a crucial step when writing term papers. Basically, scholars should read all the sources critically. In this case, the strategy allows one to understand the major concepts and ideas that relate to a research topic. Moreover, students should consider writing short notes to avoid unnecessary misunderstanding of the main messages made by authors of credible sources. Then, successful scholars take notes and revise them to ensure that they obtain the most substantial evidence that supports their research work. In turn, improving research notes involves breaking broader ideas into smaller ones and combining others to make them stronger and sensible. Therefore students should take the necessary points to support the main arguments in their term papers.    

C. Developing a Term Paper Outline

Organizing thoughts play a crucial role in preparing a quality paper. Basically, one should combine research notes obtained from scholarly sources and those gathered during brainstorming and put them into developing a term paper outline. Basically, an essay outline helps writers to connect ideas. However, a term paper outline should contain a research topic with the main thoughts and concepts needed to be covered. Besides, clear outlines have smaller ideas that relate to the main ones. In turn, the strategy allows one to see direct connections between the main ideas and leads to an organized term paper. Hence, students should follow the basic steps below to create a clear term paper outline:

• Organize notes and relevant evidence into groups of related ideas.
• Review a thesis statement to determine if it communicates the intended message.
• Identify the main points that support a working thesis statement or research hypothesis .
• Include ideas and thoughts that support the main points.
• Match supportive ideas with relevant sources obtained through research.
• Organize all the ideas to achieve a unique flow of information logically.
• Identify if some of the points presented need more research and where thoughts require development.
• Revise points and ideas to enhance the overall quality of a term paper.

D. Writing an Annotated Bibliography for a Term Paper

Successful scholars prepare annotated bibliographies that contain relevant and irrefutable research. Basically, each entry in an annotated bibliography for a term paper should include citation information with a short description and analysis. In this case, scholars should follow accepted citation styles, depending on instructions given by professors. Besides, an annotated bibliography must focus on a research topic of a term paper. The scholar should ensure that all sources remain relevant to the topic. However, one should remember that the annotated bibliography requirements may vary depending on the topic and term paper’s requirements. In turn, a useful annotated bibliography should help learners to keep track of research readings and gain a sense of a literature review . Hence, one should prepare an annotated bibliography for a term paper when conducting research.

Step 3: Start Writing a Term Paper

A. organizing a first draft of a term paper.

Drafts of the term papers help one to organize ideas in a good flow. Basically, students should use their outlines and annotated bibliographies to write the first draft of a term paper. In this case, scholars should focus on presenting all the ideas in this draft. Moreover, an appropriate draft enables one to test an outline and elaborate theories to support the central argument. In practice, good drafts resemble complete term papers. Also, good drafts should contain a title page, abstract or executive summary , introduction, body, and conclusion with a reference page.

B. Putting Everything Together

A scholar should put all the ideas together into a complete term paper. For example, learners should ensure that a written document contains a logical flow of ideas. In this case, the strategy enables students to identify some research gaps in the presented concepts of a term paper. Besides, putting everything together helps authors to identify some points that require more investigation.

C. Finding New Sources or Deleting Old Ones

Term papers must contain compelling ideas and arguments. For example, learners must review their drafts to determine if all sources provide relevant and credible evidence. In practice, scholars must change some sources that offer weak arguments. Besides, writers should remove previous scholarly sources that provide weak points of view or are irrelevant to a study since a research hypothesis may be changed during writing the first draft. Hence, one must change credible sources where necessary.

D. Altering an Outline

Learners should change their outlines of the term papers to make such pieces more substantial and compelling. For instance, one must use the first draft and new scholarly sources to make relevant changes in a term paper’s outline. Besides, the primary goal of this strategy is to strengthen study arguments and improve their clarity.

E. Creating a Working Thesis

Compelling term papers must contain well-organized thesis statements by considering research hypotheses and rationales. Basically, scholars must develop a working thesis statement that includes the claim and significant points that scholars try to make. In this case, writers should create a sentence that explains their positions taken on topics in question based on their hypotheses and rationales. Also, lecturers evaluate all the body paragraphs and how they relate to the thesis and research question. In this case, one should use a revised outline, draft, and annotated bibliography to create the working hypothesis and ensure that it meets the necessary quality.

Step 3: Wrapping It Up

A. revisions.

Outstanding term papers contain minimal or no flaws. Basically, learners must revise their drafts to remove all the mistakes. For example, some of the factors that one must consider are spelling and grammatical errors, various writing technicalities, and idea flow. In this case, revisions play a significant role in improving the term paper’s overall quality and ensuring that readers develop the motivation to evaluate all its sections. Hence, students must revise the term paper’s draft to remove unnecessary mistakes.  

Editing is an important task that helps authors to make term papers compelling. In particular, students must focus on enhancing the readability and relevance levels of a term paper. Hence, when writing a term paper, one must consider:

• Scholars should change the order of words during the editing process. In this case, the primary purpose of enhancing the term paper’s euphony is to improve the rhythm and other dynamics. Also, students should replace weak expressions to remove clichés and conversational style.
• Effective editing helps authors to enhance the brevity of statements and claims made throughout a term paper. In turn, short and concise words sound better than long or wordy statements.
• An effective editing process improves the honesty of claims made and evidence presented throughout a study work. In this case, term papers should include proven facts in each paragraph. Moreover, one should specify scholarly sources of any data used in supporting topic sentences.
• Students should edit their term papers to improve the quality of their literacy levels. For instance, one must proofread the work to remove punctuation, spelling and syntax mistakes, and typos. Also, this process requires reading a term paper several times to identify all errors and correct them accordingly.

C. Topic Sentences

Every paragraph in the body of a term paper must begin with a topic sentence. For example, learners must ensure that each section dwells on a single point related to the thesis statement. Moreover, one should reread the work to ensure that all paragraphs have the necessary opening statements.

E. Concluding Sentences

Every paragraph of a term paper should end with a concluding sentence. In this case, students should summarize covered ideas in a section. Besides, last sentences of paragraphs should include a summative claim that brings all the concepts and thoughts into a unique closure.  

F. Transitions

All ideas presented in a term paper must have a unique transitioning of ideas. For example, writers should use the necessary phrases to transition sentences and paragraphs. In particular, the approach improves the overall readability and flow of ideas in a research paper. As a rule, each paragraph’s last sentence must act as a transition to the next section. Hence, readers must find connections between all the paragraphs in a term paper.

G. Formatting

Any learning institution requires students to follow specific formatting rules. In this case, learners must follow such guidelines when writing their term papers. Also, marking rubrics are useful tools that each learner must use to format their works.

H. Peer Reviewing

Peer review is an essential step in enhancing the term paper’s quality. Basically, one should identify scholars who are familiar with a study subject to read a term paper. Also, qualified scholars help students to identify some mistakes that may undermine the term paper’s readability. Besides, peers provide positive criticism that allows students to make the necessary changes to their works.

Step 4: Writing a Final Draft of a Term Paper

The term paper’s final draft must include all the changes made during revisions, editing, formatting, and peer review. In this case, scholars should focus on submitting flawless documents that do not contain any forms of plagiarism. Besides, the final draft must capture all the aspects covered during a research study with results , discussion, recommendations, limitations, and information for further research.

Basic Outline Template of a Term Paper Format

Cover Page with a Title of a Term Paper

Abstract (150 words)

Outline (if needed)

I. Introduction

A. Relevance of research.

B. The purpose of a term paper or a discussed problem.

C. Personal reaction to a study subject.

D. Hypothesis and Rationale

E. Short descriptions of methodology and key findings.

F. Principal conclusions and thesis statement.

A. Literature Review

• Topic sentence.
• Explanation.
• Concluding sentence and transition.

B. Methodology

D. discussion.

E. Recommendations and Limitations (if needed)

III. Conclusion

A. Summary of the main points.

B. A strong response to the thesis statement.

C. A summative statement.

The outline of the term paper appears different since it contains different sections. For example, a term paper includes various subheadings that relate to the main topic. Each subheading may have several body paragraphs. However, each paragraph must contain a topic sentence, a supporting example, and a relevant explanation.

Explanations for Key Aspects of a Term Paper’s Outline Format

1. introduction part of a term paper.

The introduction must state the primary purpose of a term paper. Basically, scholars should ensure that the first part of the work acquaints readers with a problem under discussion. Besides, one must include a compelling and robust thesis statement in this section. As a rule, the introduction should not take up a large part of the entire paper. Hence, the introduction should provide an overview of the whole work in a straightforward and precise manner.

The term paper’s body should have different headings and subheadings that connect to the topic. In this case, scholars must ensure that the process of dividing a term paper into different sections enhances the clarity of the message. Moreover, the strategy should not distract readers from appreciating the intended message.

3. Conclusion

The closing paragraph should restate the thesis statement included in the introduction. Basically, students must sum up the ideas presented in all the body paragraphs. Also, the most effective strategy that one may use is to restate all the topic sentences. Besides, authors must provide a concluding statement that brings the entire work into a unique closure.

How to Write a Term Paper Proposal

A term paper proposal outlines the structure of the future work that scholars must complete. Basically, practical recommendations provide crucial elements that support the research included in the actual term paper. In this case, term proposals aim to constrict a wide area of interest into a complicated or specific topic. Moreover, writers define the intention to discover a study issue and base the decision on the need to make changes, improve the condition of the matter, or advance scholarly knowledge in the specific area of interest. In turn, one should select essential parts of a term paper and put them together in a unified format. Besides, one should briefly describe each section and tie key details to a chosen topic. Hence, a good term paper proposal should include the following parts:

• Title – A term paper should have a concise and brief title. In practice, this title should resemble that of the actual term paper.
• Objectives – Term paper proposals should state the research goals of a study. In this case, one must include the intended purpose of the research.
• Research question – An outstanding proposal must state research questions that scholars intend to answer through adequate research.
• Thesis statement – Term paper proposals should include a clear thesis statement that responds to the research question directly. In this case, a suitable thesis should be factual, clear, and subjective. Besides, one must ensure that the central claim is a verifiable statement.
• Methodology – Research proposals should state study methods used to gather and evaluate the relevant data. Moreover, students should use appropriate and proven methods to conduct their research.

Possible Writing Formats for Term Papers

Different educational institutions require learners to use writing formats when preparing term papers. For example, the most common formatting styles that one may come across include MLA 8, APA 7, Harvard, and Chicago/Turabian. In turn, each of these formats has specific guidelines that one must observe.

Term papers formatted in MLA 8 do not require a cover page unless specified. In particular, writers must include a header that contains surnames and page numbers. Moreover, one must flush these details to the right margin of the page. In turn, all in-text citations should have the author’s last name and the exact page containing the evidence used.

Term papers formatted in APA 7 should contain a title page that includes the relevant heading and identifies the student, date, and relevant institution. In this case, each page must contain a header that consists of a shortened title of the term paper and the specific page. However, the first page should have the phrase “running head” preceding the shortened title. In turn, in-text citations should contain the author’s last name and publication date.

Term paper formatted in Harvard style must contain a title page that includes the title and other details identifying the student, professor or tutor, and the relevant institution. Also, one must write the title in capital letters. Then, the header should contain the title and page number. However, one must flush these details to the right margin. As a result, in-text citations should include the author’s surname, publication date, and the page containing the relevant evidence.

4. Chicago/Turabian

Term papers formatted in Chicago/Turabian should contain a title page that has the title and identifies the author. Basically, one must capitalize all the letters in the heading. Moreover, page numbers should begin on the second page and must appear on the top right side. In turn, in-text citations should appear as footnotes that contain full bibliographic details of sources.    

Sample of Writing a Simple Term Paper

Research Topic: Is the “war on terror,” a global civil war?

Scholars develop different conceptions of the term “war on terror.” Basically, some scholars argue that the United States uses the war on terror to control the weaker nations. In this case, the current research examined whether the war on terror is a civil war. Then, a review of relevant literature was an effective method of gathering the necessary data. In turn, study results show that the United States uses the war on terror to protect citizens’ rights, which proves the alternative hypothesis to be a valid statement.

Keywords : War on terror, hypothesis, and review of literature

I. Sample of a Term Paper’s Introduction

The term war on terror became standard after the extremist attacks of September 11, 2001. Basically, President George W. Bush’s government confirmed a global campaign that involved open and secret military actions, new security lawmaking, and determinations to block extremism’s sponsoring, among other factors. In this case, the movement rallied for support from other countries, which willingly joined in the fight against terrorism. Although most scholars argue that the war on terror is an American strategy to control other countries, civil war theories prove otherwise.

Hypotheses:

H 0 – The United States uses the war on terror to control other countries.

H 1 – The United States uses the war on terror to protect the rights of its citizens.

II. Example of Body Paragraphs for a Term Paper

Different theories of civil wars prove that persistent rebel groups that engage in criminal activities. For example, some of the civil war philosophies include motivation and feasibility, ideas of rebellion, organization of the uprising, and evidence of the causes, among other factors (Kimbrough & Sheremeta, 2019). In turn, these theories prove that civil war results from the emergence and persistence of a rebel army. Moreover, motivation and feasibility theories argue that rebels with excessive power engage in terrorism. Hence, persistent rebel groups engage in terrorism as a way of addressing their interests.

A review of relevant literature is a suitable method for gathering the necessary data for the essay. According to Reale et al. (2017), a literature review is an effective method for gathering information for research papers in history and other social sciences. Therefore, reviewing scholarly sources related to civil war and terrorism will contribute to gathering the necessary data.

Standard game theory shows that economic grievances lead to terrorism. According to traditional game theory, economic dissimilarities in the community motivate some people to engage in crime (Kimbrough & Sheremeta, 2019). In this case, the less productive but healthy groups tend to participate in violence against industrious but weak groups. Besides, such conflict levels undermine fairness, equity, or coercion, depending on the analyst’s political standpoint. Hence, economic grievances motivate some groups to engage in terrorism.

Study findings show that the United States targets specific terror groups since their predatory behavior in a country or region leads to adverse economic and social outcomes. In this case, militant groups lead to redistribution through violence when productive and weak agents engage in defensive actions. Moreover, militant groups engage in activities that cause other people to reiterate. In turn, research results from such activities include reciprocated hatred that inflicts harm to innocent citizens. Hence, the United States participates in peacekeeping missions that can lower the negative impacts of such conflicts.

III. Conclusion Sample in a Term Paper

Most scholars argue that the war on terror is an American strategy to control other countries. In this case, different theories on civil wars prove that terrorism results from grievances and economic interests of some specific groups. Moreover, such activities destabilize the economic and social welfare of ordinary citizens. Thus, citizens focus on such terror groups intending to protect citizens of the affected countries. 

Kimbrough, E. O., & Sheremeta, R. M. (2019). Theories of conflict and war. Journal of Economic Behavior & Organization , 159 , 384–387. https://doi.org/10.1016/j.jebo.2019.02.007

Reale, E., Avramov, D., Canhial, K., Donovan, C., Flecha, R., Holm, P., Larkin, C., Lepori, B., Mosoni-Fried, J., Oliver, E., Primeri, E., Puigvert, L., Scharnhorst, A., Schubert, A., Soler, M., Soòs, S., Sordé, T., Travis, C., & Van Horik, R. (2017). A literature review on evaluating the scientific, social, and political impact of social sciences and humanities research. Research Evaluation , 27 (4), 298–308. https://doi.org/10.1093/reseval/rvx025

Summing Up on How to Write a Good Term Paper

Term papers refer to a research assignment completed by learners toward the end of educational semesters. In this case, students must ensure that research papers meet the necessary quality since they track and evaluate one’s knowledge. Hence, when writing a term paper, one should remember:

• rely on comprehensive research and methodological writing skills.
• include analytical and organized structures;
• present well-researched evidence that supports significant claims;
• consider various formatting strategies as required by learning institutions. 

To Learn More, Read Relevant Articles

How to write a visual analysis essay: format, outline, and example, 723 informative essay topics & ideas.

• Free Samples
• Premium Essays
• Editing Services Editing Proofreading Rewriting
• Extra Tools Essay Topic Generator Thesis Generator Citation Generator GPA Calculator Study Guides Donate Paper
• Essay Writing Help
• About Us About Us Testimonials FAQ
• Masters Term Paper
• Samples List

An term paper examples on masters is a prosaic composition of a small volume and free composition, expressing individual impressions and thoughts on a specific occasion or issue and obviously not claiming a definitive or exhaustive interpretation of the subject.

Some signs of masters term paper:

• the presence of a specific topic or question. A work devoted to the analysis of a wide range of problems in biology, by definition, cannot be performed in the genre of masters term paper topic.
• The term paper expresses individual impressions and thoughts on a specific occasion or issue, in this case, on masters and does not knowingly pretend to a definitive or exhaustive interpretation of the subject.
• As a rule, an essay suggests a new, subjectively colored word about something, such a work may have a philosophical, historical, biographical, journalistic, literary, critical, popular scientific or purely fiction character.
• in the content of an term paper samples on masters , first of all, the author’s personality is assessed - his worldview, thoughts and feelings.

The goal of an term paper in masters is to develop such skills as independent creative thinking and writing out your own thoughts.

Writing an term paper is extremely useful, because it allows the author to learn to clearly and correctly formulate thoughts, structure information, use basic concepts, highlight causal relationships, illustrate experience with relevant examples, and substantiate his conclusions.

• Studentshare

Examples List on Masters Term Paper

• TERMS & CONDITIONS
• PRIVACY POLICY
• COOKIES POLICY

• UDSIS-Students
• UDSIS-Staff
• Find It
• UD Home

Educational Technology Programs

• Performances
• Internships

Course Frameworks

• Introduction
• Grading Scale
• Course Description
• Textbooks and Computers
• Requirements
• Presentations
• 01 Understanding
• 02 Surfing and Searching
• 03 Communicating
• 04 Designing Webs
• 05 HTML Coding
• 06 Interacting with Users
• 07 JavaScript
• 09 Publishing Webs
• 10 Web Accessibility
• 11 Networking
• 12 Internet Architecture
• 13 Cybersecurity
• End-Of-Chapter Labs
• 03 Workings
• 04 Education
• 05 Cheating
• 06 Detecting
• 10 Keeping Up
• 01. Google Email
• 02. Google Calendar
• 03. Google Drive
• 04. Google Docs
• 05. Google Sites
• 06. Google Sheets
• 07. Google Slides
• 08. Google Forms
• 09. Google Meet
• 10. Google Groups
• 01 Platforms
• 02 Writing EPUBs with Pages
• 03 Writing EPUBs with MS Word
• 04 Deconstructing an EPUB
• 05 Distribution Options
• 06 Avoiding the Aggregators
• 07 Getting an ISBN (or not)
• 08 Publishing an EPUB
• 09 Troubleshooting eBook Glitches
• 10 Marketing Your eBook
• 11 Tips and Tricks
• 12 Resources
• 02 Principles
• 03 Practices
• 05 Knowledge Base
• 01 Conceptualizing
• 02 Blogging
• 03 Tweeting
• 04 Concept Mapping
• 05 Flipping the Classroom
• Discussions
• Projects Gallery
• Frameworks Gallery
• 01 Definitions
• 04 Standards
• 06 Societal Issues
• 07 Learning Theory
• 08 Internet Resources
• 09 Assessment
• 10 Planning
• 11 ePortfolio Design
• 01. What Is Multimedia?
• 02. Screen Design
• 03. Hyperlinks
• 04. Accessibility
• 05. Writing
• 06. Graphics
• 09. Teaching and Learning
• 10. Multimedia Design
• 11.Societal Issues
• 12. Multimedia Frontiers
• 13. Keeping Up
• A-Plus Project Examples
• A-Plus Term Papers
• 01 Join Apple Teacher
• 02 System Overview Badge
• 03 Pages Badge
• 04 Keynote Badge
• 05 Numbers Badge
• 06 iMovie Badge
• 07 GarageBand Badge
• 08 Get Your Certificate
• 01 Concepts
• 02 Measure App
• 03 GeoGebra 3D
• 04 Night Sky
• 05 Apollo's Moon Shot
• 06 Statue of Liberty
• 07 Berlin Wall
• 10 Reality Composer
• 01 Role of Technology
• 02 Teacher Knowledge
• 03 Personalized Learning
• 04 Equity & UDL
• 05 Collaborative Learning
• 06 Flipping or Not
• 07 Gaming & Coding
• 08 Mobile Learning
• 09 Open Learning
• 10 Sample Projects
• 01 Perspectives
• 02 Environments
• 03 Curriculum
• 04 Collaboration
• 05 Gaming and Making
• 06 List of Theories
• ePortfolio Defined
• ePortfolio Examples
• 01 Learning from iBooks
• 02 Taking Photos
• 03 Surfing with Safari
• 04 Wikipanion
• 05 Penultimate
• 06 Making Screenshots
• 07 Rotating Images
• 08 Organizing Ideas
• 09 Digital Storytelling
• 10 Dragon Dictation
• 11 Multitasking
• 12 Finding Good Apps
• 13 iTunes U
• 14 Free Books
• 15 Creating ePubs
• 16 Managing Files
• 18 GoodReader
• 19 Organizing Apps
• 20 Configuring Mail & Calendar
• 21 Securing the iPad
• 22 Searching Your iPad
• 23 Projecting the iPad
• 24 Troubleshooting
• 25 Learning More
• 01-Apple Developer
• 02-Install Xcode
• 03-Hello, World!
• 04-Sample Apps
• 05-Objective-C
• 06-Human Interface
• 07-App Design
• 08-App Store
• 02 Getting Started
• 03 Creating Channels
• 05 Embedding
• 06 Technology
• 07 Interaction
• 08 Monetization
• 09 Copyright
• 11 Censorship
• 12 References

Video Lessons

• Getting Started
• Rich Text Editing
• Pages and Subpages
• Inserting Images
• Web Page Layout
• Navigation Modes
• Making Hyperlinks
• Changing Themes

Examples of Term Papers that Got an A

Listed below are links to some term papers that got an "A" grade last year. When you compare these examples with each other, you will notice that there are three important aspects of an “A” paper. First, they are passionately written and captivating to read. Second, they have good grammar and style (following MLA, APA, or CMS style). Third, they are well documented with in-text references (in parentheses) linking their assertions to scholary articles in the list of references at the end of the paper. You will see what I mean when you follow these links to student papers that earned an “A” last year. All of these papers are copyrighted by their authors. Please respect these copyrights.

• Aisha-McCormick-Digital-Marketing
• Allie-Modica-Effectiveness
• Allison-Winters-Technology-And-Learning-Styles
• Amanda-Yanez-Social-Media-Advertising
• Amanda-Warren-Social-Media-Likes
• Austin-Abbruzzesi-Computer-Science-Classes-in-High-School-Curricula
• Ben-Rohe-MOOCs
• Brianna-Patrizio-Gender-Equality
• Collette-Small-Social-Media
• Danielle-Piha-CyberBullying
• Danielle-Ragno-LinkedIn
• David-Palgon-Facebook-and-Employers
• Donna-Muchio-Self-Esteem
• Ebenezer-Riverson-3D-Printing
• Elizabeth-Hansen-WII-Physical-Therapy
• Faith-Lumpkin-LinkedIn
• Giselle-Malenchek-Digital-Piracy
• Jessica-Morris-Healthy-Lifestyles
• Katy-Snyder-Social-Media-Advertisers
• Jillian-Loeffler-Social-Media-in-Your-Job-Search
• Lauren-Vandaniker-Technology-Impact-On-Nursing-Practice
• Lisa-Plumley-Multimedia-Young-Children
• Madison-Gamble-Technology-As-A-Means-For-Therapeutic-Modalities
• Maria-Medved-Hospitality-Industry
• Lisa-Tossey-POV-Cameras-Education
• Paul-Page-Copyright-IP-Public-Good
• Sarah-Ibarguen-Twitter
• Taylor-Soave-Social-Media-Classroom
• Tim-Ware-Social-Media-News-Outlet

• School of Education  •   Willard Hall Education Building  •   Newark, DE 19716  •   USA Undergraduate phone: 302-831-8695  •  Fax: 302-831-4110 Graduate phone: 302-831-1165  •  Fax: 302-831-4421 Director's Office: 302-831-3178  •  Fax: 302-831-6039
• Legal Notices

• Buy Essays Online
• Buy admission essay
• College Essay Writing Service
• Economics essay writing service
• Law essay writing service
• College paper writing service
• Research paper writing service
• Term paper writing service
• PowerPoint presentation homework help
• Write my book report
• Assignment writing service
• Lab report writing service
• Programming assignment help
• Statistics assignment help
• Article Critique
• Annotated bibliography writing service
• Case study writing service
• Coursework writing service
• Dissertation Writing Services
• Speech writing service
• Thesis writing help

Term Paper Outline

Your term paper outline is your reliable compass throughout the writing process. Here, you arrange all the points that you are going to discuss in your term paper. Mostly you do it for yourself.

It will always help you stay focused and stick to the main points in your paper. However, a tutor, teacher, or professor may ask you to submit your working-term paper outline before you start writing. He or she will check the ways that you are going to develop your thesis and can suggest some improvement areas.

You may revise and restructure this version of your outline once you come across new material or find new ideas to be included in your term paper.

Don't make haste writing your paper. Think carefully about your topic and main points as well as evaluate the material you have found. Subdivide all the relevant material into groups and then name each group. These names will serve as headings and subheadings in your outline.

Our paper writing service would like to remind you that every term paper, regardless of the subject, has the following sections:

• Introduction or purpose of the paper. This opening part suggests acquainting the reader with the problem and stating the thesis.
• Heading 1: History of the Problem. You may include past attempts at solutions.
• Heading 2: Extent of the Problem. Who is affected? What impact has it had?
• Heading 3: Effects of the Problem.
• Heading 4: Possible Future Solutions.
• Conclusion : Sums up the points made in the term paper and gives a strong answer to the thesis.

Term Paper Writing

You haven’t got yourself admitted in the college for just a mediocre degree to get mediocre jobs in your life, have you? Sure, even getting a C or D would get you that degree but why bother settling for less when you can get an A+?

As a student, you must have familiarity with the term papers. A lot of students think that great term paper writing requires Shakespeare level writing skill. However, in reality, above average writing skill is enough to write a striking term paper – all you need to be is strategic and organized.

So, do you want to know the secret recipe of writing a remarkable term paper that can help you get the ever-eluding A+? Just go through the whole guide, and you will get the gist.

What is a Term Paper?

Let’s start with the term paper definition. By definition, a term paper is a type of research-based writing assignment that a student has to submit to his or her teacher at the end of an academic term. Typically, a student tries to discuss elaborately on a topic that was assigned to him or her. The topic could be an event description, a case study, a concept, or an argument. It is mandatory that the paper has to be unique – plagiarism is not allowed.

An average term paper is about several pages in length. There is no fixed length, and often the corresponding teacher has the authority to impose structure and length for writing. In such a case, a student must adhere to the teacher's instructions.

However, the commonly accepted length of such paper is about 5,000 words. With the single line spacing, single column, 12pt font, and 1inch margined general term paper format; it should be about 15 pages. We will talk about the structure and formatting style later.

Differences between Term Paper and Research Paper

Students often confused between these two different types of papers. The first difference between them is the time frame – a term paper has to be submitted by the end of the semester or a term while a research paper may need months even years to complete.

Even the task was given out in a different timeline. Generally, the teacher asks the students to work on their research paper at the beginning of the academic semester. A term paper, on the other hand, is tasked somewhere in the middle of the semester.

Another major difference between them is the purpose. The purpose of a research paper is to find a viable solution to a problem while a term paper is the reflection of knowledge of student on a particular topic – in common cases, a description.

Moreover, a research paper has a hypothesis, to begin with, by the end, it either supports the hypothesis or rejects it with feasible data. A term paper is much simpler in nature and tries to support an existing thesis paper in most cases. That’s why a research paper has a significantly longer length.

Despite a research paper being more crucial, it doesn't affect a student’s final grade while a term paper strongly controls the grades.

The Basics of Term Paper Format: What are the Parts of a Term Paper?

A term paper has a fixed format, but it differs from course to course. For example, a Computer Science course will have a different format of writing when compared to an Architecture course. You must ask your teacher about how to outline a paper – he or she can guide you the best.

Just check out the most common format that could help you outlining a term paper.

How to Write a Term Paper?

There are a few simple ideas you can follow in crafting up a brilliant piece of writing. Here are some effective ideas for you.

Be prepared for LONG research hours

Research is a must for writing a term paper. Some students make a mistake by starting to write right away. This is a rookie mistake. Although it seems that there is no progress as not a single word is being written, research would make your writing phase much faster and fluent. Moreover, you could come up with a better approach and idea if you thoroughly research topic first. Think of it as the investment for writing the paper.

Create an outline first

Creating an outline prior to writing gives you a sense of control. You can how much time it would take to complete the whole piece and which part would take more time. At the same time, you would render a smoother reading experience, and everything would be in some kind of chronological order.

Come up with a compelling introduction

"Dawn shows the day!" If you have an impelling introduction, the reader will get more interested in reading through and eventually favor you and your grades. If you lose the grip in the introduction, no matter how good your content is, your reader will give it average feedback.

Avoid fluff words

Fluff words are attention killers. Especially, when you are writing formal academic writing, the reader would seek constructive information all along the writing. You don't need to unnecessarily blabber about anything as long as you are talking about something relevant.

Conclude with the ROCC method

The conclusion is just as much important as the introduction. A great way of writing an impactful conclusion is following the ROCC method. ROCC stands for Restating your standpoint, having One vital and strong gist, Concluding tone, and leaving a Clincher for a reader to think about.

Select a citation style

Proper formatting or following a citation style is a must while writing a research paper or a term paper. Following the MLA or the APA format is a wise decision. However, make sure you are sticking to either MLA or APA, a mix between these two looks really shabby.

Make sure to proofread

Yes, you have worked really hard writing up the whole term paper, and you aren't not feeling proofreading the whole paper. Although it sounds really painful to proofread, you might correct A LOT OF misspellings and simple grammatical mistakes. You don't want your grades gone to ashtray just because you feel lazy, do you?

Term Paper Topics

A term paper is known to be one of the most important papers in your study. That's why picking the right topic for your paper is critical. Moreover, a lot of students fail their term papers because of the wrong formatting. So, our team decided to help with the first step in writing a term paper and give you some tips on how to cite it correctly.

50 original topics from our term paper writing service 

All these topics are creative ideas from our writing team. We are tired of writing student's orders on abortion and other common topics, so we decided to share our ideas with you. Your professor will be impressed by your creativity. These topics are completely original and should help you to boost your grades. Moreover, if you stuck with such a topic, our academic writing service is ready to help you!

Whether you are looking for a college term paper or the university one – these picks will apply to both. We divided our list into five main categories, like education, environmental issues, family issues, social issues, and the political ones. So, enjoy the list, and don't forget to ask for help from our team!

Term paper topics on education 

If you got stuck with the education topic for your term paper, we are here to help you. Our top picks will definitely surprise your supervisor and give you more chances to get the higher grade. These topics can be reworked or widened. So here are our ten topics our team created for you:

• Pros and cons of distance learning in college.
• The consequences of plagiarism in a student's paper.
• Why should students have continuous sex education?
• Reasons for installing metal detectors in schools.
• How do social networks influence school life?
• How can modern technologies change the way we learn?
• Bullying at school and college and how to take action?
• Why do we need a multilingual education?
• Is it worth investing in a child's education?
• How to solve sex discrimination issues in schools?

 Term paper topics on environment 

Environmental issues topics are vast, they are covering a lot of disciplines in natural sciences fields. It is hard to pick the right one because of so many choices out here. So, our online term paper writing team decided to take a step and pick the top ten grade-boosters for you:

• Why do a lot of people thinking that global warming is a hoax?
• Why is recycling highly important for the environment?
• How to overcome water lacking issues and help everyone on the Earth?
• The advantages and disadvantages of nuclear energy?
• Wildfires conservation – are there more environment-friendly alternatives?
• Is it possible to manage overpopulation?
• Can we predict hurricane impacts and get prepared?
• What are the types of alternative energy can save Earth natural resources?
• Should we film more movies about environmental issues?
• Way to reduce smog in the megalopolises?

Term paper topics on family issues 

Family is the basis of our society. Yet, not all of them are healthy. This topic set we created is devoted to family issues, and you should try to focus on the solutions. Our team wrote a lot of term papers on this topic for students studying social sciences, so we know how to pick the perfect one:

• The consequences of battered woman syndrome.
• Child abuse at home and possible resolutions.
• Divorce reasons and US statistics: why happy couples are breaking up?
• Domestic abuse and possible legal actions.
• Children in divorced families: how do parents living separately influence life?
• Toxic relationships and the ways to break-up with a toxic partner.
• How to overcome the loss of a family member: the psychology of children and adults?
• Generation gap: start understanding your beloved ones.
• Mentally challenged family member: how to live normally and care of your beloved one.
• Ageism: why an older family member is not always right and how to influence the situation.

Term paper topics on Politics 

Politics vector changes each year. Politicians are making right or wrong decisions changing the relationship inside and outside of the country. You might agree or disagree with them, and this term paper is a chance to share your thoughts. Here are our top picks for this set:

• Controversial political decisions that saved millions of people
• Totalitarianism: is it now exist? How are people fighting with it?
• How to solve the corruption issues in politics?
• Are elections important for politics?
• What is the difference between the US and the UK political systems? Which one is better?
• What is the ideal political system and how to build it?
• What is the role of public relations and media in politics?
• What causes revolutions? Is it a political failure?
• How does politics influence cultural development?
• What is a democracy and how it is different from socialism?

Term paper topics on Social issues 

Social issues related to society. Cultural, moral and ethical aspects influence them. So, while picking the topic for your paper, you should be thinking about the cultural aspect of an issue. Our team has collected ten amazing social issues topics you may consider for your next big thing:

• What are the reasons for country-wide strikes and protests?
• Worldwide flash mobs and why people participate?
• Can virtual reality and communities substitute the real world? What are the consequences?
• The ways to fight racism in the country.
• Why should the Church and the state remain separate?
• Women and trans people rights: how to avoid discrimination?
• Why may we need a death penalty?
• Humanitarian missions: are they effective?
• How beauty standards influence mental health?
• HIV/AIDS people social isolation and how to influence the issue?

APA formatting – how to format your term paper right

If you are studying psychology in college or university, your papers should be styled following APA guidelines. Here is a brief must have a list for this complicated style:

• The text should be typed on A4 white paper with the dimensions of 8.5" x 11".
• You need to use a 12pt Times New Roman font since it is easy to read.
• Use double spacing throughout the paper.
• Margins of 1" should be set on all sides, i.e. at the left, right, top and bottom of your paper.
• Set paragraph indention to 1/2 inch, it can be easily done with the Tab button in your word processor.
• You need to create a first page header which consists of the page number and as well as capitalized running head you have on your title page.

MLA formatting rules for a term paper

According to our  term paper writing service  experts MLA is the most chosen paper formatting directive. That's why it's highly important to understand its rules and follow all the guidelines. So here are several things about this formatting style for your paper:

• You should use 12-point Times New Roman font.
• Your paper should be double-spaced. Make sure that there no extra spaces or single spacing throughout your paper.
• At the upper left corner of your first page, you need to list your name, instructor's name, class, and the date. All should be at the new row.
• You should have a one-inch margin on each side of your paper.
• Your last name and page number should be added to the upper right corner of each page, including the first one.
• You should have a title of your paper which is centered and appear under your heading details. The title is formatted like the rest of your paper. Never underline your title, don't use bold or italics, as well as quotation marks.
• Indent each paragraph to the right. Indentation should be set to 1/2 inch.
• Align all your text to the left.

Now, you have everything needed for term paper writing. If you need help, don't hesitate to contact our team. Our writing experts are ready to help you!

If you are determined and focused enough, you can write great term papers all by yourselves. You cannot neglect your term paper as it carries a significant amount of weight that can either give you a great grade or destroy it! So, we hope you know what is a term paper now and how you can write a great one.

Related Services:

• Paper writing service
• Term paper help
• Dissertation writing services
• Amendments $30
• Title page $5
• Bibliography $5
• E-mail delivery $10
• Formatting $10
• Plagiarism check of each writer
• True Quality
• Guaranteed Privacy
• Satisfaction guarantee
• On time delivery
• No hidden charges
• 24/7 Live Support

Unique offer for a first-time customer

Send us your email and receive it

Done! Check your email for the discount

Check Promotions and Spam folders if you can`t find the letter.

We use cookies. What does it mean? OK

Find theses and term papers for your studies

Access a variety of term papers, bachelor theses and master theses, why should you use it.

Let’s help each other and encourage science

Scientific writing

Learn how other students write their research papers

Latest sources

Get the latest scientific sources and speed up your literature research

Mutual support

Become part of our community and get better grades

Topic-specific theses & term papers

Find a variety of academic papers

Search for topics and find term papers, bachelor theses and master theses.

Share your final- and term papers

Share your knowledge with other students and become part of our community.

Save valuable time

Save time on your research, improve your writing skills and get better grades.

From students, for students

Bachelor theses, master theses, term papers.

Search for related research papers

Type in related words to your topic and start browsing other student research papers

What our students say

Best experience!

“Awesome site. I wish I had it already years ago for my first term papers. It helped me a lot.”

Taylor Stewart

“StudyVent is an excellent source of knowledge. It helps me to get new ideas on papers and saves much time on the process.“

Jessica Holder

Highly recommended!!

“Easy to manage and it provides me a starting point on my paper!“

Rachel Brown

• Term Paper Topics
• Custom Term Papers

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

• View all journals
• My Account Login
• Explore content
• About the journal
• Publish with us
• Sign up for alerts
• Open access
• Published: 10 April 2024

FOXO1 is a master regulator of memory programming in CAR T cells

• Alexander E. Doan 1   na1 ,
• Katherine P. Mueller   ORCID: orcid.org/0000-0002-1873-3638 2 , 3 , 4   na1 ,
• Andy Y. Chen   ORCID: orcid.org/0000-0002-5018-355X 5 , 6 , 7   na1 ,
• Geoffrey T. Rouin 2 , 3 , 4 ,
• Yingshi Chen   ORCID: orcid.org/0000-0002-5391-5103 2 , 3 , 4 ,
• Bence Daniel   ORCID: orcid.org/0000-0002-2410-8767 5 , 7 , 8 , 9   nAff18 ,
• John Lattin 1 ,
• Martina Markovska 2 , 3 , 4 ,
• Brett Mozarsky 2 , 3 , 4 ,
• Jose Arias-Umana   ORCID: orcid.org/0000-0002-7154-4780 2 , 3 , 4 ,
• Robert Hapke 2 , 3 , 4 ,
• In-Young Jung 10 ,
• Alice Wang   ORCID: orcid.org/0000-0002-1789-8561 2 ,
• Peng Xu 1 ,
• Dorota Klysz   ORCID: orcid.org/0000-0002-6733-6110 1 ,
• Gabrielle Zuern 2 , 3 , 4 ,
• Malek Bashti   ORCID: orcid.org/0000-0003-0398-3367 1 ,
• Patrick J. Quinn   ORCID: orcid.org/0000-0002-4406-727X 1 ,
• Zhuang Miao 9 ,
• Katalin Sandor   ORCID: orcid.org/0000-0003-2892-4865 5 , 7 ,
• Wenxi Zhang 5 , 7 ,
• Gregory M. Chen 4 , 11 ,
• Faith Ryu 2 , 3 , 4 ,
• Meghan Logun   ORCID: orcid.org/0000-0002-7274-811X 4 , 12 ,
• Junior Hall 2 , 13 , 14 ,
• Kai Tan 2 , 13 , 14 ,
• Stephan A. Grupp 2 , 13 , 14 ,
• Susan E. McClory 2 , 13 ,
• Caleb A. Lareau   ORCID: orcid.org/0000-0003-4179-4807 5 , 7 , 15 ,
• Joseph A. Fraietta   ORCID: orcid.org/0000-0001-7900-8993 4 , 10 , 11 , 14 ,
• Elena Sotillo   ORCID: orcid.org/0000-0002-3993-1932 1 ,
• Ansuman T. Satpathy   ORCID: orcid.org/0000-0002-5167-537X 5 , 7 , 15 ,
• Crystal L. Mackall   ORCID: orcid.org/0000-0001-6323-4304 1 , 15 , 16 , 17   na2 &
• Evan W. Weber   ORCID: orcid.org/0000-0001-5039-2899 2 , 3 , 4 , 13 , 14 , 15   na2  

Nature ( 2024 ) Cite this article

11k Accesses

278 Altmetric

Metrics details

• Cancer immunotherapy
• Immunotherapy

A major limitation of chimeric antigen receptor (CAR) T cell therapies is the poor persistence of these cells in vivo 1 . The expression of memory-associated genes in CAR T cells is linked to their long-term persistence in patients and clinical efficacy 2 , 3 , 4 , 5 , 6 , suggesting that memory programs may underpin durable CAR T cell function. Here we show that the transcription factor FOXO1 is responsible for promoting memory and restraining exhaustion in human CAR T cells. Pharmacological inhibition or gene editing of endogenous FOXO1 diminished the expression of memory-associated genes, promoted an exhaustion-like phenotype and impaired the antitumour activity of CAR T cells. Overexpression of FOXO1 induced a gene-expression program consistent with T cell memory and increased chromatin accessibility at FOXO1-binding motifs. CAR T cells that overexpressed FOXO1 retained their function, memory potential and metabolic fitness in settings of chronic stimulation, and exhibited enhanced persistence and tumour control in vivo. By contrast, overexpression of TCF1 (encoded by TCF7 ) did not enforce canonical memory programs or enhance the potency of CAR T cells. Notably, FOXO1 activity correlated with positive clinical outcomes of patients treated with CAR T cells or tumour-infiltrating lymphocytes, underscoring the clinical relevance of FOXO1 in cancer immunotherapy. Our results show that overexpressing FOXO1 can increase the antitumour activity of human CAR T cells, and highlight memory reprogramming as a broadly applicable approach for optimizing therapeutic T cell states.

Similar content being viewed by others

cBAF complex components and MYC cooperate early in CD8+ T cell fate

Ao Guo, Hongling Huang, … Douglas R. Green

Stem-like exhausted and memory CD8+ T cells in cancer

Thomas Gebhardt, Simone L. Park & Ian A. Parish

Low-dose decitabine priming endows CAR T cells with enhanced and persistent antitumour potential via epigenetic reprogramming

Yao Wang, Chuan Tong, … Weidong Han

More than 50% of patients who respond to CAR T cell therapies eventually relapse, and CAR T cells that target solid tumours have been largely ineffective 1 . The expression of memory T cell genes in patient CAR T cells is associated with durable persistence and disease control 2 , 3 , 4 , 5 , 6 , but the transcription factors that drive CAR T memory programs have not been identified. We previously showed 7 that providing rest to exhausted CAR T cells through transiently inhibiting CAR signalling promoted a memory-like phenotype and increased chromatin accessibility at motifs bound by the memory transcription factors TCF1 and FOXO1, raising the prospect that these transcription factors mediate memory programming in CAR T cells. Consistent with this notion, expression of TCF7 (which encodes TCF1) broadly correlates with responses to CAR T cell 2 , 5 , tumour-infiltrating lymphocyte (TIL) 8 and checkpoint blockade 9 , 10 therapies. In addition, FOXO1 directly regulates the expression of TCF7 and other canonical memory genes 11 , 12 and promotes the formation of central memory T cells in mice 12 , 13 , 14 .

Several groups have shown that pharmacological inhibition of AKT, a negative regulator of FOXO1, confers an early memory phenotype in human CAR T cells and TILs 15 , 16 , 17 , suggesting that FOXO1 also promotes memory in human T cells. To test the hypothesis that FOXO1 is required for memory programming and antitumour function in human CAR T cells, we performed phenotypic and functional experiments using CD19.28ζ or CD19.BBζ CAR T cells cultured in the presence of a selective FOXO1 small-molecule inhibitor 18 (FOXO1 i ) (Extended Data Fig. 1 ). FOXO1 i reduced the expansion and viability of CAR T cells, the frequency of CD8 + cells and the expression of memory-associated markers (CD62L, IL-7Rα and TCF1) in a dose-dependent manner, and concomitantly upregulated markers of short-lived effector or exhausted T cells (CD39, TIM-3 and TOX) (Extended Data Fig. 1b–e ).

We corroborated these data by using CRISPR–Cas9 to knock out FOXO1 (FOXO1 KO ) (Fig. 1a and Extended Data Fig. 2a ). FOXO1 KO CAR T cells showed a similar reduction in expansion and CD8 + frequency, diminished memory-associated markers and increased exhaustion-associated markers as compared with  AAVS1 -edited control CAR T cells (Fig. 1b,c , Extended Data Fig. 2b–f and Supplementary Fig. 1 ). Because FOXO1 KO cells exhibited uniformly low CD62L surface expression, we used CD62L as a surrogate marker for FOXO1 editing by magnetically purifying CD62L lo FOXO1 KO cells for bulk RNA sequencing (RNA-seq) (Extended Data Fig. 2g,h ). FOXO1 KO cells upregulated activation- and exhaustion-associated genes ( TOX , NR4A1 , FOS and CD69 ), downregulated memory and FOXO1 target genes ( IL7R and CCR7 ) and exhibited less naive-like and more exhausted gene-expression signatures (Fig. 1d,e and Extended Data Fig. 2i ).

a – g , CRISPR–Cas9 gene editing of AAVS1 (AAVS1) or FOXO1 (FOXO1 KO ) in CD19.BBζ CAR T cells. a – c , Flow cytometric analysis of FOXO1 knockout efficiency ( a ), percentage of CAR + CD8 + cells at day 14 ( b ) and memory- and exhaustion-associated markers in CAR + CD8 + cells ( c ). Shaded areas in a represent gates used in phenotypic analyses. One representative donor is shown in a  and  c ( n  = 6 donors). d , Volcano plot of DEGs in CD62L lo FOXO1 KO versus AAVS1 (Bonferroni-adjusted P  < 0.05 with absolute log 2 -transformed fold change (abs(log 2 (FC)) > 0.5). e , GSVA using T cell gene signatures 55 . f , Cytokine secretion in response to Nalm6 leukaemia cells from one representative donor ( n  = 4 donors). g , Stress test Nalm6 xenograft model. Top, schematic. Bottom, survival curves of Nalm6-engrafted mice treated with mock T cells or gene-edited CD19.BBζ cells. Data show two donors tested in two independent experiments ( n  = 8 or 9 mice per group). Data in  d  and e  include n  = 3 donors. h – n , CAR T cells overexpressing truncated NGFR (tNGFR), TCF1-P2A-tNGFR (TCF1 OE ) or FOXO1-P2A-tNGFR (FOXO1 OE ). h , Flow cytometric analysis of FOXO1, TCF1 and CD19.28ζ expression from one representative donor ( n  = 8 donors). FMO, fluorescence minus one. i – k , Serial restimulation of CD19.BBζ cells with Nalm6. CD8 + CAR T cell expansion ( i ) and flow cytometric analysis of memory- and exhaustion-associated markers ( j , k ). j , Mean ± s.e.m. of normalized mean fluorescence intensity (MFI) ( n  = 2 or 3 donors). k , One representative donor ( n  = 4 donors). l , HA.28ζ cytokine secretion (day 13) in response to 143B osteosarcoma cells from one representative donor ( n  = 4 donors). m , n , HA.28ζ seahorse analysis (day 13) ( n  = 2 donors). m , Oxygen consumption rate (OCR) (mean ± s.d. of 11 technical replicates from one representative donor). Oligo, oligomycin; R+A, rotenone and antimycin. n , Spare respiratory capacity. Data in  f , l , n are mean ± s.d. of three technical replicates. Statistical comparisons were performed using paired two-tailed Student’s t -test ( b , e ), two-sided Welch’s t -test ( f ), log-rank Mantel–Cox test ( g ) and repeated-measures one-way ANOVA with Geisser–Greenhouse correction ( j ) or one-way ANOVA with Dunnett’s test ( l , n ) .

Source Data

FOXO1 i and FOXO1 KO cells also exhibited attenuated killing and/or cytokine secretion after tumour challenge (Fig  1f and Extended Data Fig. 1f,g ), consistent with a model in which FOXO1 restrains exhaustion and/or terminal differentiation in human T cells, similar to reports in mice 14 , 19 , 20 , 21 , 22 . We corroborated these results using an in vitro CAR T cell exhaustion model (HA.28ζ CAR), in which antigen-independent tonic CAR signalling induces features of exhaustion within approximately one week 7 , 23 . Knockout of FOXO1 in HA.28ζ cells accelerated the manifestation of exhaustion markers and dysfunction (Extended Data Fig. 2j,k ). We next modelled chronic antigen stimulation in vivo by infusing a sub-therapeutic dose of CD19.BBζ cells into leukaemia-bearing mice 7 , 24 . Knockout of FOXO1 significantly reduced CAR T cell tumour control and survival (Fig. 1g ). These observations show that endogenous FOXO1 promotes memory and is required for optimal antitumour function of CAR T cells.

FOXO1 overexpression preserves a memory phenotype

Among the genes induced by FOXO1 is TCF7 , which has been broadly implicated in memory programming, stemness and antitumor activity in human and mouse T cells 2 , 5 , 8 , 10 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 . Thus, we sought to determine whether the overexpression of FOXO1 and/or TCF1 could enhance the function of human CAR T cells. Human T cells were co-transduced with a retrovirus expressing a CAR and a second virus expressing truncated NGFR (tNGFR) as a control or a bicistronic vector containing tNGFR and either TCF1 (TCF1 OE ) or FOXO1 (FOXO1 OE ) (Extended Data Fig. 3a ). This approach enabled high levels of transcription factor overexpression and equivalent CAR expression across conditions (Fig. 1h ). Notably, CD19.BBζ cells expressing FOXO1 OE , but not TCF1 OE , exhibited increased baseline expression of memory-associated surface markers and transcription factors, including endogenous TCF1 (refs. 12 , 13 ) (Extended Data Fig. 3b,c ).

TCF1 OE and FOXO1 OE cells that were serially rechallenged with Nalm6 leukaemia both exhibited enhanced cytokine secretion compared with controls (Extended Data Fig. 3d ), but only FOXO1 OE increased CD8 proliferation and memory marker expression while suppressing the levels of TOX (Fig. 1i–k and Extended Data Fig. 3e ). By contrast, TCF1 OE increased the expression of TOX and CD39 relative to tNGFR controls, consistent with a more exhausted or effector-like phenotype (Fig. 1j and Extended Data Fig. 3e ). We corroborated these results in cells expressing the tonic signalling HA.28ζ CAR, in which both TCF1 OE and FOXO1 OE cells showed enhanced function, but only FOXO1 OE promoted a memory-like surface phenotype (Fig. 1l and Extended Data Fig. 3f–h ).

Because the metabolism of memory T cells favours oxidative phosphorylation (OXPHOS) relative to glycolysis, we used Seahorse to assess whether transcription factor overexpression induces memory-like metabolic profiles. FOXO1 OE and TCF1 OE showed increased OXPHOS and superior metabolic fitness compared with tNGFR controls. The degree of FOXO1 OE -mediated metabolic reprogramming was more marked in exhausted HA.28ζ cells (Fig. 1m,n ) compared with those expressing CD19.28ζ (Extended Data Fig. 3i,j ), consistent with the notion that FOXO1 OE counteracts the exhaustion program.

FOXO1 OE promotes a memory-like gene signature

We hypothesized that FOXO1 and TCF1 induce disparate gene-expression programs because overexpression of each endowed CAR T cells with distinct cell-surface phenotypes and functionality (Fig. 1 ). Therefore, we performed bulk RNA-seq on purified CD4 + or CD8 + FOXO1 OE and TCF1 OE T cells expressing HA.28ζ to model settings of chronic antigen stimulation. Principal component analysis (PCA) showed that FOXO1 OE and TCF1 OE CAR T cells clustered separately from tNGFR and had a greater number of unique differentially expressed genes (DEGs) than shared genes (Fig. 2a,b and Extended Data Fig. 4a–c ). PCA also showed that transcription factor overexpression was a stronger driver of differential gene expression than CD4 + or CD8 + cell identity (Extended Data Fig. 4b ), confirming that FOXO1 OE and TCF1 OE promote divergent gene-expression programs in both subsets.

a – l , Bulk RNA-seq ( a – f ) and ATAC-seq ( g – l ) in tNGFR + CD8 + HA.28ζ CAR T cells ( n  = 3 donors). a , RNA-seq PCA. b , Venn diagram showing unique and shared DEGs in TCF1 OE and FOXO1 OE compared with tNGFR (Bonferroni-adjusted P  < 0.05 with abs(log 2 (FC) > 0.5). c , GSVA of DEGs using naive (T N ), T pex and exhausted (T ex ) T cell signatures 55 . Centre line represents mean score. d , Heat map and hierarchical clustering of DEGs. Genes of interest are highlighted. The colour bar shows normalized z -scores for each DEG. e , f , GO term analyses showing curated lists of the top upregulated and downregulated processes in FOXO1 OE ( e ) and TCF1 OE ( f ) versus tNGFR (Benjamini–Hochberg-adjusted P ). g , ATAC-seq PCA. h , Number of differentially accessible peaks compared with tNGFR ( P  < 0.05 with abs(log 2 FC) > 0.5). i , GSVA of differentially accessible peaks using an early T ex cell epigenetic signature 9 . Centre line represents mean score. j , Chromatin accessibility tracks for the IL7R , KLF3 , TOX and FASLG loci, for one representative donor. k , l , Rank-ordered plots of differentially accessible transcription factor (TF)-binding motifs in FOXO1 OE ( k ) and TCF1 OE ( l ) versus tNGFR. ZF, zinc-finger. Statistical comparisons were performed using DESeq2 ( b , d , h , k , l ), one-sided hypergeometric test ( e , f ) and repeated-measures one-way ANOVA with Dunnett’s test ( c , i ).

Gene set variation analysis (GSVA) showed that FOXO1 OE promoted a naive-like and less terminally exhausted gene signature (Fig. 2c ). Consistent with these data, HA.28ζ FOXO1 OE cells upregulated genes associated with memory ( SELL , IL7R , LEF1 and TCF7 ) and downregulated those associated with exhaustion ( TOX , HAVCR2 , ENTPD1 and CD244 ) (Fig. 2d and Extended Data Fig. 4d,e ). Despite the fact that previous literature has implicated FOXO1 in regulatory T (T reg ) cell biology 34 , 35 , FOXO1 OE did not enforce a T reg gene signature (Extended Data Fig. 4f ). Gene ontology (GO) and ingenuity pathway analysis (IPA) showed that FOXO1 OE promoted autophagy, cellular catabolism and naive-associated transcription factor gene-expression networks ( TCF7 and LEF1 ) and diminished effector transcription factor networks ( ID2 , PRDM1 and TBX21 ) (Fig. 2e,f and Extended Data Fig. 4g,h ). By contrast, TCF1 OE cells exhibited high expression of exhaustion-associated transcription factors of the NR4A family, a progenitor exhausted T (T pex ) cell-like gene signature (Fig. 2c ), and were enriched in effector gene-expression pathways (for example, cell–cell adhesion, T cell activation and cytokine production) (Fig. 2d,f and Extended Data Fig. 4d,e,g ). Similar results were obtained in CD19.28ζ cells (Extended Data Fig. 4i–k ); however, FOXO1 OE resulted in a greater number of DEGs in tonic signalling HA.28ζ CAR T cells compared with those expressing CD19.28ζ, indicating more marked transcriptional reprogramming by FOXO1 during chronic stimulation.

TCF1 and FOXO1 are considered pioneer factors owing to their ability to directly bind to condensed chromatin and recruit chromatin remodelling machinery 36 , 37 . To test whether TCF1 OE and/or FOXO1 OE induce chromatin remodelling, we performed a bulk assay for transposase-accessible chromatin with sequencing (ATAC-seq) in TCF1 OE and FOXO1 OE CAR T cells (Supplementary Fig. 2 ). PCA confirmed that both transcription factors promoted global changes to chromatin accessibility compared with tNGFR controls (Fig. 2g and Extended Data Fig. 5a ). This effect was most evident in tonically signalling HA.28ζ cells, in which FOXO1 OE clustered separately from tNGFR and TCF1 groups and showed more differentially accessible peaks (around 5,600; P  < 0.05) compared with TCF1 OE cells (around 3,000) (Fig. 2g,h ). Most of the differentially accessible peaks in FOXO1 OE were open, consistent with the ability of FOXO1 ability to perturb core histone–DNA contacts 37 .

HA.28ζ FOXO1 OE cells showed increased accessibility at FOXO1 target gene loci ( IL7R and KLF3 ), reduced accessibility at exhaustion-associated loci ( TOX and FASLG ) and a decreased exhaustion-like epigenetic signature compared with tNGFR cells (Fig. 2i,j ), consistent with transcriptomic data. Of note, DNA-binding motifs for transcription factors of the forkhead box and HMG-box families were the top-ranked differentially accessible motifs in FOXO1 OE and TCF1 OE cells, respectively (Fig. 2k,l and Extended Data Fig. 5b,c ), supporting a model in which overexpressed FOXO1 and TCF1 induce local chromatin remodelling. Paradoxically, FOXO1 OE cells also showed increased accessibility at transcription factor motifs associated with effector function (for example, b-ZIP and NF-κB p65) (Extended Data Fig. 5d,e ).

These data show that FOXO1 OE induces memory and naive-like gene-expression programs during chronic stimulation, whereas TCF1 OE promotes a T pex -like program, consistent with the role identified for TCF1 in chronic infection and cancer 25 , 26 , 38 , 39 . In addition, FOXO1 OE induces a unique epigenetic state that supports effector function while maintaining memory programming.

FOXO1 OE  enhances CAR T function against leukaemia

Because FOXO1 OE was effective at promoting memory (Fig. 1 ), we hypothesized that further increasing the activity of FOXO1 might endow CAR T cells with a more stable memory phenotype. We generated a humanized version of a nuclear-restricted variant of FOXO1 (FOXO1 3A ), which is insensitive to AKT-mediated nuclear export 19 (Extended Data Fig. 6a–c and Supplementary Fig. 3 ). FOXO1 3A increased the surface expression of FOXO1 target genes to a similar extent to FOXO1 OE (Extended Data Fig. 6d,e ). However, FOXO1 3A expression induced a divergent transcriptomic profile that was de-enriched in T cell activation genes and led to blunted in vitro cytokine secretion and cytotoxicity compared with FOXO1 OE (Extended Data Fig. 6f–i ). These observations raised the prospect that excessive nuclear FOXO1 activity might promote a stable memory phenotype and oppose effector function 21 .

To assess function in a protracted model in which memory programming might be important for sustained antitumor activity, we used a stress test xenograft model in which leukaemia-bearing mice received a sub-therapeutic dose of CD19.28ζ (Fig. 3a ) or CD19.BBζ (Extended Data Fig. 7a ) CAR T cells. FOXO1 OE markedly enhanced the tumour control of CAR T cells compared with tNGFR, whereas TCF1 OE showed no benefit (Fig. 3a and Extended Data Fig. 7a,b ). Similar results were obtained in a curative Nalm6 model, in which FOXO1 OE cells exhibited increased expansion and persistence compared with TCF1 OE and tNGFR cells (Fig. 3b and Extended Data Fig. 7c–e ). FOXO1 3A provided a modest survival advantage compared with tNGFR, but FOXO1 3A cells exhibited delayed expansion and reduced levels of tumour control compared with FOXO1 OE cells (Fig. 3a–c ), consistent with the notion that FOXO1 3A partially opposes effector function. To assess the recall response to secondary antigen challenge—a hallmark feature of memory T cells 40 —we rechallenged nearly cured mice with a high dose of Nalm6 (Fig. 3b,c and Extended Data Fig. 7c–e ). Only FOXO1 OE cells re-expanded after rechallenge and conferred a survival advantage, showing that FOXO1 OE endows CAR T cells with superior in vivo effector- and memory-like functions compared with tNGFR, TCF1 OE or FOXO1 3A .

a , Subcurative doses of 0.1 × 10 6 –0.2 × 10 6 tNGFR + CD19.28ζ cells were infused into Nalm6-bearing mice seven days after engraftment. Schematic (top) and survival curve (bottom) are shown; n  = 9-10 mice per group. b – d , Curative doses of 1 × 10 6 tNGFR + CD19.28ζ cells were infused into Nalm6-bearing mice seven days after engraftment. Mice were rechallenged with 10 × 10 6 CD19 + or CD19 − Nalm6 on day 21 after CAR T cell infusion ( n  = 2 donors tested in 2 independent experiments). b , Rechallenge Nalm6 model. Schematic (top) and quantification (bottom) of circulating human CD45 + CAR T cells. Mean ± s.e.m. of n  = 3–7 mice per group from one representative donor. c , Survival curve after rechallenge ( n  = 3–8 mice per group pooled from 2 donors). d – f , CD19.28ζ cells overexpressing tNGFR or FOXO1 OE were gene-edited to knock out AAVS1 (control; AAVS1 ) or TCF7 ( TCF7 KO ). d , RNA-seq PCA. e , Volcano plots of DEGs; n  = 3 donors (Bonferroni-adjusted P  < 0.05 with abs(logFC) > 0.5). f , Stress test Nalm6 model. tNGFR + CD19.28ζ cells (0.6 × 10 6 cells) were infused into Nalm6-bearing mice seven days after engraftment. Survival curve is shown ( n  = 8–10 mice per group). a , c , f show pooled data from two donors tested in two independent experiments. Statistical comparisons were performed using log-rank Mantel–Cox test ( a , c , f ) and DESeq2 ( e ). NS, not significant.

TCF7 is not required for FOXO1 OE reprogramming

To investigate the mechanism by which FOXO1 OE reprograms CAR T cells and increases in vivo antitumour activity, we generated a variant of FOXO1 with lower-affinity DNA binding (FOXO1 DBD ) 41 . FOXO1 DBD showed a modest reduction in DNA binding, and its expression in CAR T cells perturbed FOXO1-mediated transcriptional and epigenetic reprogramming (Extended Data Fig. 8a–d ). Mice that received CD19.28ζ FOXO1 DBD cells showed reduced survival in a Nalm6 leukaemia stress test model compared to those that were infused with FOXO1 OE cells (Extended Data Fig. 8e ), indicating that FOXO1 OE DNA binding is crucial for augmented antitumour activity.

The FOXO1 target gene, TCF7 ), is highly upregulated in FOXO1 OE cells (Extended Data Figs. 3c and 4d ). Although TCF1 OE did not increase the potency of CAR T cells, we reasoned that high endogenous levels of TCF7 and expression kinetics in FOXO1 OE could be mechanistically important for FOXO1 OE reprogramming. Notably, knockout of TCF7 in the context of FOXO1 OE had negligible effects on FOXO1 OE transcriptional reprogramming and in vivo antitumour activity (Fig. 3d–f and Extended Data Fig. 8f ). Thus, FOXO1 OE reprogramming requires DNA binding but not transcription of the memory-associated transcription factor and target gene, TCF7 .

FOXO1 OE enhances CAR T function in solid tumours

To determine whether FOXO1 was also capable of increasing the activity of CAR T cells against solid tumours, we infused tNGFR or FOXO1 OE HER2.BBζ CAR T cells into 143B osteosarcoma-bearing NSG mice. Consistent with leukaemia models, FOXO1 OE cells showed durable antitumour activity and persistence (Fig. 4a–e and Extended Data Fig. 9a–d ). Tumour-infiltrating FOXO1 OE cells exhibited transcriptomic reprogramming, were enriched in gene signatures associated with T cell killing, effector function and tissue residence, and showed negligible differences in human T reg signatures 42 , 43 (Fig. 4f–h and Extended Data Fig. 9e–i ). Of note, intratumoral FOXO1 OE cells did not have a canonical memory-like phenotype but were enriched in a FOXO1 OE transcriptomic signature derived from bulk RNA-seq studies (Fig. 4h ), suggesting that exogenous FOXO1 remains active in the tumour microenvironment.

A total of 5 × 10 6 mock or tNGFR + Her2.BBζ CAR T cells expressing tNGFR or FOXO1 OE were infused into 143B-bearing mice three days after engraftment. a , b , Tumour measurements over time ( a ) and on day 25–29 ( b ). One FOXO1 OE mouse has been omitted in b owing to tumour-independent death before day 25. Data were pooled from three donors tested in three independent experiments ( n  = 11–18 mice per group). c – e , Analysis of day-29 CAR TILs. c , Total CAR TILs ( n  = 13 mice per group). d , Ratio of CD8 + to CD4 + CAR TILs. One representative donor ( n  = 10 mice per group). e , CAR TIL IL-2 and IFNγ secretion after ex vivo stimulation with 143B ( n  = 13 mice per group). Data in c – e were pooled from two donors tested in two independent experiments. f – h , Single-cell RNA-seq on day-29 CAR TILs. Cells were sorted and pooled from n  = 5 mice per group from one donor. f , Left, uniform manifold approximation and projection (UMAP) of CAR TILs. Eleven clusters were identified with k -nearest neighbours clustering, and were annotated manually (middle). Right, sample distribution by cluster. T eff , T effector cell; T RM , tissue resident memory T cell. g , Sample distribution within the UMAP. h , T eff , T RM and FOXO1 OE -associated transcriptional signatures. Long dashed lines represent the mean and short dashed lines represent the top and bottom quartiles. Data in b – e are mean ± s.e.m. Statistical comparisons were performed using two-tailed Student’s t -test ( b – e ) and two-sided Wilcoxon rank-sum test ( h ).

Together, these data show that FOXO1 OE increases the in vivo expansion, persistence and tumour control of CAR T cells in a TCF7 -independent manner, whereas TCF1 OE provides no measurable benefit. FOXO1 OE -mediated enhancements are dependent on DNA binding and nuclear export, which suggests that tuning or signal regulation mediated by nuclear shuttling is important for effective FOXO1-mediated memory programming.

FOXO1 activity correlates with response to T cell therapies

FOXO1 target genes, including TCF7 , were enriched in pre-infusion CAR T cells that mediate clinical responses in patients 2 , 5 (Extended Data Fig. 10a,b ), raising the possibility that endogenous FOXO1 activity might predict potent antitumour activity in clinical CAR T products. Paradoxically, however, FOXO1 transcript levels in pre-infusion CD19.BBζ cells were not associated with response to therapy or survival in adults with chronic lymphocytic leukaemia (CLL) (Fig. 5a and Extended Data Fig. 10c ). Because FOXO1 is regulated mainly post-translationally rather than transcriptionally 15 , we hypothesized that the activity of FOXO1 could be better approximated by the aggregate expression of FOXO1 target genes. We therefore identified a FOXO1 ‘regulon’ consisting of 41 overlapping DEGs that were downregulated in FOXO1 KO cells and upregulated in FOXO1 OE cells (Fig. 5b ). The FOXO1 regulon included putative FOXO1 target genes (for example, SELL and KLF3 ), but was made up largely of genes that have not previously been associated with memory programming (Supplementary Table 1 ). In contrast to FOXO1 transcript, the FOXO1 regulon was significantly enriched in pre-infusion CAR T cells from patients with CLL who exhibited complete or partial responses with transformed disease, and was associated with in vivo CAR T cell expansion and overall survival (Fig. 5c,d and Extended Data Fig. 10d ). TCF7 did not reach statistical significance in FOXO1 KO experiments and was therefore not included in the FOXO1 regulon; however, regulon score significantly correlated with the TCF7 transcript in patient CAR T cells, suggesting that the regulon is an accurate readout for FOXO1 transcriptional activity (Fig. 5e ).

a – e , Single-sample gene set enrichment analysis (ssGSEA) on RNA-seq from pre-infusion, CAR-stimulated CTL019 cells from patients with CLL 2 (complete responder (CR), n  = 5; partial responder with transformed disease (PR TD ), n  = 3; partial responder (PR), n  = 5; non-responder (NR), n  = 21). a , FOXO1 ssGSEA for patient outcomes (left) and overall survival (right). b , The FOXO1 regulon was generated using FOXO1 KO and FOXO1 OE bulk RNA-seq data and then applied to published datasets 2 , 5 ; n  = 3 donors. c , FOXO1 regulon ssGSEA (data from ref. 2 ) for patient outcomes (left) and overall survival (right). d , Least squares regression (dark line) of FOXO1 regulon score and peak CAR T cell expansion. e , Simple linear regression (dark line) of TCF7 expression and FOXO1 regulon score. Dark lines in a , c represent patient survival curves and shaded areas in a , c , e represent 95% confidence intervals. Dots in d , e represent individual samples (blue, CR/PR TD ; grey, NR/PR). f , FOXO1 regulon ssGSEA for pre-manufactured effector T cells from paediatric patients with B-ALL with durable (six or more months of B cell aplasia (BCA); n  = 33 patients) or short (less than six months of BCA; n  = 27 patients) CAR T cell persistence 5 . g , An epigenetic signature derived from FOXO1 OE ATAC-seq was applied to pre-manufactured T cell single-cell ATAC-seq data from paediatric patients 5 . Data show FOXO1 OE epigenetic signature scores for patients with durable (patient 52, n  = 616 cells; patient 54, n  = 2,959 cells) and short (patient 38, n  = 2,093 cells; patient 66, n  = 2,355 cells) CAR T cell persistence. h , GSEA using FOXO1 OE DEGs and DEGs derived from CD39 − CD69 − TILs from adult patients with melanoma 8 . ES, enrichment score. Violin plots in a , c , f , g show minima and maxima; solid lines represent the mean and long dashed lines represent the top and bottom quartiles. Statistical comparisons were performed using two-tailed Mann–Whitney test ( a , left; c , left; f ), log-rank Mantel–Cox test ( a , right; c , right), Spearman correlation ( d , e ), two-sided Wald test ( g ) and two-sided Kolmogorov–Smirnov test ( h ).

The FOXO1 regulon was also enriched in pre-manufactured effector T cells from children with B cell acute lymphoblastic leukaemia (B-ALL) who exhibited durable CAR T cell persistence 5 (Fig. 5f ), supporting the notion that FOXO1 activity broadly correlates with the efficacy of CAR T cells. Because both FOXO1 and TCF1 mediate chromatin remodelling 36 , 37 , 44 , 45 , 46 (Fig. 2 ), we next used epigenetic signatures derived from our ATAC-seq analyses to interrogate single-cell ATAC-seq data from paediatric CAR T cells 5 . Consistent with FOXO1 regulon transcriptomic data, the FOXO1 OE epigenetic signature was significantly enriched in patient T cells that were associated with durable persistence, whereas the TCF1 OE signature was not (Fig. 5g and Extended Data Fig. 10e ). Finally, FOXO1 OE DEGs were enriched in stem-like CD39 − CD69 − TILs that were highly predictive of the response to TIL therapy in adult patients with melanoma 8 , whereas TCF1 OE DEGs were de-enriched (Fig. 5h and Extended Data Fig. 10f ).

In this study, we tested the hypothesis that overexpressing memory-associated transcription factors could reprogram CAR T cells to durably persist and maintain antitumor activity. We focused our efforts on FOXO1, on the basis of studies that have implicated this transcription factor in memory programming 12 , 13 , 14 , 19 , 20 , 21 , 46 , 47 , 48 , 49 , 50 , 51 and our previous work in which we showed that exhaustion reversal and memory programming were associated with enhanced chromatin accessibility at FOXO1-binding motifs 7 . FOXO1 overexpression induced memory gene-expression programs and chromatin remodelling, mitigated exhaustion and substantially improved persistence and antitumour function in four distinct xenograft models. Its effect was independent of CAR binder, co-stimulatory domain and tumour type, highlighting the broad applicability of this pro-memory program across CAR T cell products.

There is a vast body of literature describing the role of FOXO1 in promoting T cell memory and persistence in mice 12 , 13 , 14 , 19 , 20 , 21 , 46 , 47 , 48 , 49 , 50 , 51 ; however, FOXO1 biology in human T cells remains poorly understood. Because the activity of FOXO1 is regulated at the post-translational level rather than through changes in transcription and is therefore hidden in RNA-seq data, the role of FOXO1 in cancer immunology and immunotherapy is likely to have been considerably underappreciated. Our study is the first, to our knowledge, to show that endogenous FOXO1 is required for memory gene expression and optimal antitumour function in engineered human T cells, which is consistent with the effects of Foxo1 knockout in mouse models of acute and chronic infection 14 , 19 , 20 . We further show that endogenous FOXO1 restrains exhaustion in human T cells, because deleting FOXO1 induced an exhaustion-like phenotype and CAR T cell dysfunction.

Notably, FOXO1 activity in pre-infusion CAR T cells and TILs strongly correlated with clinical responses, underscoring the importance of FOXO1 in T-cell-based cancer immunotherapies. Paradoxically, expression of a nuclear-restricted variant (FOXO1 3A ) altered FOXO1 reprogramming and attenuated the antitumour function of CAR T cells, supporting the notion that optimal FOXO1 activity involves intermittent and/or context-dependent regulation. Indeed, others have shown that transient expression of FOXO1 3A can induce partial memory reprogramming in human CAR T cells without impairing effector function 15 , 52 , 53 . Further work is needed to determine how FOXO1 expression levels and kinetics affect the function of CAR T cells and whether FOXO1 is relevant in other therapeutic modalities, such as immune checkpoint blockade.

We also interrogated TCF1, a transcription factor that defines stem-like or memory T cell populations that exhibit an increased capacity to respond to immune checkpoint blockade 2 , 5 , 8 , 10 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 . Of note, overexpressing TCF1 did not enforce memory gene-expression programs or enhance antitumour activity in vivo, which contradicts reports in mice 27 , 28 . Instead, TCF1 OE cells exhibited a gene-expression signature associated with T pex cells, and manifested functional hallmarks of exhaustion during chronic stimulation, consistent with other studies 39 , 54 . Thus, our results raise the possibility that constitutive TCF1 overexpression skews human engineered T cells towards a more exhausted or T pex  cell-like state, and/or that TCF7 -expressing T pex cells do not have a substantial role in CAR T cell responses.

An alternative interpretation posits that FOXO1, rather than TCF1, is mainly responsible for endowing tumour-reactive T cells with a stem-like or progenitor phenotype, and that TCF7 expression is merely a readout for FOXO1 activity. Indeed, deletion of endogenous TCF7 in FOXO1 OE did not affect FOXO1-mediated transcriptional reprogramming or augmented antitumour function in vivo. Surface markers and transcription factors that are often co-expressed in TCF7 + cells are FOXO1 target genes 29 , and our empiric FOXO1 regulon significantly correlated with TCF7 expression and clinical responses in samples of CAR T cells from patients, further supporting this notion. Conditional deletion of Foxo1 in mature mouse T cells diminished the frequency of Tcf7 -expressing T pex cells 14 , suggesting that FOXO1 might promote cell states that are normally associated with high levels of Tcf7 expression. Future mechanistic studies are warranted to determine the precise roles of FOXO1 and TCF1 in human engineered and non-engineered T cells during cancer immunotherapy.

In summary, we show that FOXO1-driven transcriptional and epigenetic programs are associated with engineered and non-engineered T cells that expand, persist and promote clinical responses in patients with cancer. Overexpression of FOXO1 increases the activity of CAR T cells through memory reprogramming, and TCF1 is insufficient to induce CAR T cell memory and persistence. Our results suggest that FOXO1 represents a major therapeutic axis that can be exploited to improve the efficacy of T-cell-based cancer immunotherapies.

Primary human T cells

For experiments completed at Stanford, buffy coats from anonymous, consenting healthy donors were obtained from the Stanford University Blood Center under an University Institutional Review Board-exempt protocol or obtained from a human peripheral blood leukopak (STEMCELL Technologies). CD3 + cells were isolated using the RosetteSep Human T Cell Enrichment Kit, Lymphoprep density gradient medium and SepMate-50 tubes according to the manufacturer’s protocol (STEMCELL Technologies). For experiments completed at the Children’s Hospital of Philadelphia (CHOP), purified CD3 + healthy donor T cells were obtained from the University of Pennsylvania Human Immunology Core. All purified T cells were cryopreserved in CryoStor CS10 medium (STEMCELL Technologies).

Cell lines were obtained from ATCC and stably transduced to express markers as follows: 143B osteosarcoma cells express GFP and firefly luciferase with or without CD19, Nalm6 B-ALL cells express GFP and firefly luciferase with or without GD2. Single-cell clones were chosen for high antigen expression. The 143B and Nalm6 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) and RPMI 1640, respectively, and both were supplemented with 10% fetal bovine serum (FBS), 10 mM HEPES and 1× penicillin–streptomycin–glutamate (Gibco). Nalm6 and 143B cell lines and engineered versions of these cell lines were previously authenticated via STR fingerprinting prior to their use in this study. HEK293 cells were originally obtained from the National Cancer Institute. Cells were frequently tested for mycoplasma using the Lonza MycoAlert Mycoplasma Detection kit.

Design of CAR and transcription factor constructs

The CAR constructs used in this study include CD19.28ζ, CD19.BBζ, anti-GD2 HA.28ζ and Her2.BBζ. Codon-optimized TCF1, FOXO1 or FOXO1 3A sequences and a P2A ribosomal skip sequence were generated as Gene Blocks by IDT and constructed in MSGV retroviral vectors. The tNGFR-only construct does not contain a P2A ribosomal skip sequence. The FOXO1 DBD construct was generated by two-step mutagenic NEBuilder HiFi DNA Assembly (New England BioLabs). All plasmids were amplified by transformation into Stellar Competent Escherichia coli (Takara Bio), and sequences were validated by sequencing (Elim Biopharmaceuticals).

Retrovirus production

To generate retrovirus, ten million 293GP cells were plated on a 15-cm BioCoat poly- d -lysine cell culture plate (Corning) and fed with 20 ml of DMEM supplemented with 10% FBS, 10 mM HEPES and 1× penicillin–streptomycin–glutamate (Gibco) 24 h before transfection. Transfection was performed by mixing a room-temperature solution of 3.4 ml Opti-MEM (Gibco) + 135 μl Lipofectamine 2000 (Invitrogen) (solution 1) with a second solution of 3.4 ml Opti-MEM + 11 μg RD114 packaging plasmid DNA + 22 μg MSGV retroviral plasmid of interest (solution 2) by slow dropwise addition of solution 2 to solution 1. The combined solution 1 and 2 mixture was incubated for 30 min at room temperature, after which the medium was replaced on 293GP cells, and 6.5 ml of the combined solution was added to the plates in a slow, dropwise manner. The next day, the culture medium was replaced on 293GP cells. At 48 h after transfection, the viral supernatant was collected from the cells and the culture medium was replaced; supernatant collection was repeated at 72 h. At each step, the supernatant was spun down to remove cells and debris, and frozen at −80 °C for future use.

T cell activation and culture

T cells were thawed in warm water after removal from liquid nitrogen and then washed with T cell medium (AIM-V (Gibco) supplemented with 5% FBS, 10 mM HEPES, 1× penicillin–streptomycin–glutamate and 100 U ml −1 recombinant human IL-2 (Peprotech) or RPMI (Gibco) supplemented with 10% FBS, 10 mM HEPES, 1× penicillin–streptomycin–glutamate and 100 U ml −1 recombinant human IL-2). Human T-Expander αCD3/CD28 Dynabeads (Gibco) were washed and added to T cells at a volume of 30 μl resuspended beads per million T cells. T cells and beads were then resuspended at a concentration of 500,000 T cells per ml in T cell medium (day 0 for all assays). Forty-eight and 72 hours after activation, T cells were transduced (see ‘Retroviral transduction’). Ninety-six hours after activation, beads were removed by magnetic separation using a DynaMag column (Invitrogen). T cells were fed with fresh T cell medium every 48–72 h and were maintained at a density of 0.5 ×10 6 cells per ml after feeding. For FOXO1 i experiments, T cells were provided with fresh complete T cell medium and vehicle control (dimethyl sulfoxide; DMSO) or AS1842856 (EMD Millipore) every 2–3 days from days 4 to 15 after activation.

Retroviral transduction

T cells were transduced with retrovirus on days 2 and 3 after activation for all experiments. In brief, 12- or 24-well, non-tissue-culture-treated plates were coated with 1 ml or 500 μl, respectively, of 25 μg ml −1 Retronectin (Takara) in PBS and placed at 4 °C overnight. The next day, plates were washed with PBS then blocked with 2% bovine serum albumin (BSA) + PBS for 10 min. Retroviral supernatants were added and plates were centrifuged at 32 °C for 2 h at 2,500 g . Viral supernatants were subsequently removed and T cells were added to each virus-coated well at a density of 1 × 10 6  T cells per well for 12-well plates and 0.5 × 10 6  T cells per well for 24-well plates.

Cell selection

tNGFR isolations were performed using either Miltenyi MACS sorting or STEMCELL EasySep sorting unless otherwise stated. For Miltenyi MACS sorting, cells were resuspended in FACS buffer and stained with biotin anti-human CD271 (tNGFR) antibody (BioLegend). Cells were washed with PBS, 0.5% BSA and 2 mM EDTA (MACS buffer), resuspended in MACS buffer and mixed with Streptavidin MicroBeads (Miltenyi), then washed again with MACS buffer and passed through an LS Column for positive selection inside a MACS separator (Miltenyi). For STEMCELL EasySep sorting, cells were isolated using the manufacturer’s protocol for the EasySep Human CD271 Positive Selection Kit II (STEMCELL Technologies) with an EasyEights EasySep Magnet (STEMCELL Technologies). After isolation, cells were immediately mixed with warm complete T cell medium, counted and resuspended at 500,000 per ml.

For RNA-seq experiments on FOXO1 KO cells, CD62L lo CAR + cells were isolated by negative selection, first by staining cells with anti-CD62L-PE and then by following the EasySep PE Positive Selection Kit II protocol according to the manufacturer’s instructions (STEMCELL Technologies). For RNA-seq and ATAC-seq experiments on tNGFR, TCF1 OE and FOXO1 OE cells, CD8 + tNGFR + CAR T cells were isolated before sequencing using the EasySep Human CD8+ T Cell Isolation Kit (STEMCELL Technologies). For in vivo analysis of tumour-infiltrating CAR T cells, CD45 + T cells were isolated from tumours using the EasySep Release Human CD45 Positive Selection Kit (STEMCELL Technologies) according to the manufacturer’s instructions.

CRISPR–Cas9 gene editing

To interrogate the role of endogenous FOXO1 in CAR T cell function, CRISPR–Cas9 was used to delete a sequence directly upstream of the FOXO1 DNA-binding domain. On day 4 after activation, retrovirally transduced CAR T cells were removed from activation beads by magnetic separation. Twenty-microlitre reactions were prepared by resuspending one million CAR T cells in P3 buffer immediately before electroporation with the P3 Primary Cell 4D Nucleofector Kit (Lonza). Ribonucleoproteins were prepared by complexing 0.15 ng of sgRNA targeting FOXO1 or AAVS1 (Synthego) with 5 µg Alt-R S.p. Cas9 Nuclease (IDT, 1081058) before adding the cell suspension to each reaction. For AAVS1 edits, a previously validated sgRNA sequence (5′-GGGGCCACUAGGGACAGGAU-3′) was used. For FOXO1 , two separate sgRNAs were used in tandem, at equal concentrations (5′-UUGCGCGGCUGCCCCGCGAG-3′ and 5′-GAGCUUGCUGGAGGAGAGCG-3′). For TCF7 gene editing, we used a previously validated sgRNA 56 (5′-UCAGGGAGUAGAAGCCAGAG-3′) for bulk RNA-seq experiments performed at CHOP. A separate sgRNA (5′-UUUUCCAGGCCUGAAGGCCC-3′) was designed and validated at Stanford, and used for in vivo experiments. The reaction was pulsed with the EH115 program on a Lonza 4D Nucleofector. Cells were recovered immediately in 260 µl of warm complete AIM-V medium supplemented with 500 U ml −1 IL-2 in round-bottom 96-well plates and expanded into 1 ml fresh medium within 24 h. Cells were maintained at 0.5 × 10 6  cells per ml to 1.0 × 10 6 cells per ml in well plates until day 14–16 for functional and phenotypic characterization. On days 14–16, knockout efficiency was determined by intracellular transcription factor staining (Cell Signaling, 58223) followed by flow cytometry.

Flow cytometry

CAR T cells were washed twice in FACS buffer (PBS + 2% FBS) and stained with fluorophore-conjugated surface antibodies for 30 min on ice. Cells were washed twice with FACS buffer before analysis. Intracellular stains were performed with the same initial surface stain, after which cells were fixed, permeabilized and stained using the FoxP3 Transcription Factor Staining Buffer Set according to the manufacturer’s protocol (eBioscience). Anti-human FOXO1 (clone C29H4) and anti-human TCF1 (C36D9) antibodies were purchased from Cell Signaling. The 1A7 anti-14G2a idiotype antibody used to detect the HA CAR was obtained from the NCI and conjugated using the Dylight 650 antibody labelling kit (Thermo Fisher Scientific). The anti-FMC63 idiotype antibody was manufactured by GenScript and fluorescently conjugated using the Dylight 650 antibody labelling kit. Cell-surface antibodies were used at a 1:100 dilution during staining, with the exception of anti-14g2a and anti-FMC63, which were used at a 1:1,000 dilution. Intracellular antibodies were used at a 1:50 dilution and live/dead staining was used at a 1:1,000 dilution. Cells were analysed with either a BD Fortessa running FACS Diva software, or a Cytek Aurora using SpectroFlo v.3.1.0. Downstream analyses were performed using Cytek SpectroFlo v.3.1.0 and FlowJo v.10.8.1 Software. All reagents are listed in Supplementary Table 2 . A representative gating strategy for FOXO1 KO and FOXO1 OE experiments is shown in Supplementary Fig. 1 . In experiments in which we stained for Annexin V, cells were gated on all singlets, excluding debris but not excluding dead or dying T cells. For MFI quantification, background subtraction was performed using either unstained or FMO samples. The MFI quantification in Extended Data Fig. 1e was not background subtracted owing to negative MFI values in some control samples.

Cytokine secretion assays

A total of 5 × 10 4 CAR T cells were co-cultured with 5 ×10 4 tumour cells in 200 μl of complete T cell medium (AIM-V or RPMI) without IL-2 in a 96-well plate, all in triplicate. Twenty-four hours after co-culture, culture supernatants were collected, diluted 20- to 100-fold and analysed for IL-2 and IFNγ using ELISA MAX kits (BioLegend) and Nunc Maxisorp 96-well ELISA plates (Thermo Fisher Scientific). Absorbance readings were collected on a Tecan Spark plate reader or a BioTek Synergy H1 running Gen5 v.2.00.18. For FOXO1 i assays, the co-culture medium included concentrations of AS1842856 that were used during T cell expansion.

IncuCyte killing assay

A total of 5 × 10 4 GFP + tumour cells and T cells corresponding to a 1:1, 1:2, 1:4, 1:8 and/or 1:16 effector:target ratios were co-cultured in 300 μl of T cell medium without IL-2 in 96-well flat-bottom plates. Plates were imaged at 10× zoom with 4–9 images per well every 2–4 h for 96 h using the IncuCyte ZOOM S3 Live-Cell analysis system (Essen BioScience/Sartorius). The total integrated GFP intensity per well or total GFP area (μm 2 per well) were used to analyse the expansion or contraction of Nalm6 or 143B cells, respectively. All GFP intensity and area values were normalized to the first imaging time point ( t  = 0). For FOXO1 i assays, the co-culture medium included concentrations of AS1842856 that were used during T cell expansion.

Repeat stimulation assay

CAR T cells were activated and transduced, and tNGFR + cells were isolated as described above. Cells were cultured in AIM-V with IL-2 until day-14 ‘pre-stim’ assays, including flow cytometry, cytokine secretion and IncuCyte as described above. On day 14, co-cultures were set up comprising 5 × 10 T cells and 2 × 10 6 Nalm6 tumour cells suspended in AIM-V without IL-2 at a final concentration of 5 × 10 5 total cells per ml. Co-cultures were fed with 5 ml of AIM-V without IL-2 on day 3 of culture. On day 3 of the repeat stimulation co-culture, CAR T cells were again assayed by cytokine secretion, IncuCyte killing assay and flow cytometry as described above. This process was repeated for a total of four co-cultures such that the cytokine and IncuCyte assays were set up for four serial stimulations on days 14, 17, 20 and 23 on cells that had been stimulated with Nalm6 tumour zero, one, two and three previous times, respectively, for a total of four serial stimulations by the end of the experiment. Cells were analysed by flow cytometry on day 7 of co-culture, such that T cells were co-cultured with tumour on days 14, 17, 20 and 23 and analysed on days 21, 24, 27 and 30, respectively.

Seahorse assay

Metabolic analyses were performed using Seahorse Bioscience Analyzer XFe96. In brief, 0.2 × 10 6 cells were resuspended in extracellular flux assay medium supplemented with 11 mM glucose, 2 mM glutamine and 1 mM sodium pyruvate, and plated on a Cell-Tak (Corning)-coated microplate allowing the adhesion of CAR T cells. Mitochondrial activity and glycolytic parameters were measured by the oxygen consumption rate (OCR) (pmol min −1 ) and extracellular acidification rate (ECAR) (mpH min −1 ), respectively, with the use of real-time injections of oligomycin (1.5 M), carbonyl cyanide ptrifluoromethoxyphenylhydrazone (FCCP; 0.5 M) and rotenone and antimycin (both at 0.5 M). Respiratory parameters were calculated according to the manufacturer’s instructions (Seahorse Bioscience). Reagent sources are listed in Supplementary Table 2 .

Immunoblotting

Chromatin-bound and soluble proteins were separated as previously described 23 . In brief, cytoskeletal (CSK) buffer was prepared using 100 mM NaCl, 300 mM sucrose, 3 mM MgCl 2 , 10 mM PIPES (pH 6.8), 0.1% IGEPAL CA-630, 4 µg ml −1 aprotinin, 10 µg ml −1 leupeptin, 4 µg ml −1 pepstatin and 2 mM PMSF. After washing with ice-cold PBS, cell pellets were lysed with CSK buffer for 20 min on ice. Samples were centrifuged at 1,500 g for 5 min and the soluble fraction was separated and cleared by centrifugation at 15,870 g for 10 min. The protein concentration of the soluble fraction was determined by DC protein assay (Bio-Rad, 5000116). The remaining pellet containing the chromatin-bound fraction was washed twice with CSK buffer, centrifuging at 1,500 g for 5 min. Chromatin-bound proteins were resuspended in CSK buffer and 1× Pierce Reducing Sample Buffer (Thermo Fisher Scientific, 39000) and boiled for 5 min for solubilization. The soluble fraction was supplemented with Pierce Reducing Sample Buffer to achieve 1× and boiled for 5 min. For immunoblotting, equal amounts of soluble and chromatin-bound fraction for each sample were analysed by SDS–polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes (Bio-Rad, 1704158). Membranes were blocked for 30 min in 5% milk in TBST (1× Tris-buffered saline containing 0.1% Tween-20). After washing with TBST, membranes were incubated with anti-FOXO1 antibody (1:1,000; Cell Signaling, 2880, clone C29H4) overnight at 4 °C. Next, membranes were washed with TBST and incubated with anti-mouse (1:10,000, Cell Signaling, 7074) or anti-rabbit (1:10,000, Cell Signaling, 7076) IgG conjugated to horseradish peroxidase for 1 h at room temperature. Membranes were visualized using Clarity Western ECL Substrate (Bio-Rad, 1705060) and the ChemiDoc Imaging System and Image Lab Touch Software v.3.0 (Bio-Rad). After visualization, membranes were stripped using a mild stripping buffer (1.5% glycine, 0.1% SDS, 1% Tween-20, pH 2.2). The previous steps were repeated for detection of soluble (1:5,000 GAPDH; Cell Signaling, 97166, clone D4C6R) and chromatin-bound (1:1,000 Lamin A; Cell Signaling, 86846, clone 133A2) fraction loading controls. Densitometry analyses were performed using Fiji v.2.14.0/1.5 f.

Mouse xenograft models

NOD/SCID/ Il2rg −/ − (NSG) mice were bred, housed and treated under Stanford University APLAC- or CHOP ACUP-approved protocols. Six-to-eight-week-old mice were healthy, immunocompromised, drug- and test-naive and unused in other procedures. Mice were housed at the Stanford Veterinary Service Center (VSC) or CHOP Department of Veterinary Services (DVR) in a barrier facility with a 12-h light–dark cycle, and mice were kept at a temperature of 20–23 °C (CHOP) or 20–26 °C (Stanford) with humidity ranging from 30–70%. Five mice were housed in each cage in aerated racks with ample bedding, food and water. For mice that became sick, solid feeds were switched to liquid feeds to facilitate eating. Mice were monitored daily by trained VSC and DVR staff under the supervision of a veterinarian who reported excess morbidity immediately and/or euthanized mice for humane reasons. Mice were euthanized if end-point criteria were met, which included 143B tumour sizes exceeding 1.2 cm or Nalm6 bioluminescence greater than 5 × 10 11 photons per second, or if evidence of extensive disease occurred (for example, inability to ambulate, groom or eat, cachexia, excessive loss of fur, hunched posture or other signs of disability); whichever came first. Tumour injection sites were chosen so as not to interfere with the mouse’s normal body functions, such as ambulation, eating, drinking, defecation and/or urination. In Nalm6-bearing mice, 2 × 10 5 to 1 × 10 7 cells in 100–200 μl of sterile PBS were engrafted by tail vein injection (TVI). In 143B osteosarcoma models, 1 × 10 6 to 3 × 10 6 cells in 100 μl sterile PBS were engrafted by intramuscular injection into the flank. Mice were randomized prior to CAR T cell infusion to ensure equal tumour burden across groups. CAR T cells were engrafted by TVI at doses and schedules noted in the main text. Nalm6 engraftment, expansion and clearance were measured by intraperitoneal injection of luciferin and subsequent imaging by a Spectrum IVIS bioluminescence imager and quantified using Living Image software v.4.7.3 (Perkin Elmer), or by a Lago X imager and quantified using Aura software v.4.0.7 (Spectral Instruments Imaging), all under isoflurane anaesthesia. The 143B tumour size was monitored by caliper measurements. Tumor and T cell injections were performed by technicians who were blinded to treatments and expected outcomes.

Mouse tissue analyses

Peripheral blood was sampled from live, isoflurane-anaesthetized mice by retro-orbital blood collection. Fifty microlitres of blood was labelled with surface antibodies, lysed using FACS Lysing Solution (BD) and quantified using CountBright Absolute Counting Beads (Thermo Fisher Scientific), then analysed on a BD Fortessa cytometer. For phenotypic analysis of spleen and tumours, mice were euthanized and tissues were mechanically dissociated and washed twice in PBS. Spleens were placed in a 6-cm Petri dish and filtered through a sterile 70-µm cell strainer. Tumours were mechanically and chemically dissociated with Collagenase IV and DNAse in HBSS and incubated at 37 °C with shaking for 30 min. Cells were mashed through a sterile 70-µm cell strainer before washing with PBS. Cells from both spleens and tumours were spun down at 450 g for 5 min at 4 °C, then treated with ACK lysis buffer for 3 min on ice. Cell suspensions were washed twice with PBS and CAR T cells were isolated by positive selection using the EasySep Release Human CD45 Positive Selection Kit. Cells were stained for markers of interest and analysed on a Cytek Aurora using SpectroFlo Software 3.1.0.

Bulk RNA-seq

A total of 0.5 × 10 6 –1 × 10 6  T cells were pelleted by centrifugation and flash-frozen. Pellets were thawed on ice and processed using either an RNEasy Plus Mini Kit or an AllPrep DNA/RNA Micro Kit (for simultaneous DNA and RNA isolation) (QIAGEN) according to the manufacturer’s instructions. Total RNA was quantified using either a Qubit Fluorometer or a DeNovix DS-11 FX Spectrophotometer/Fluorometer and sequenced using a 150 bp paired-end read length and around 50 million read pairs per sample (Novogene).

Bulk RNA-seq processing and analysis

We processed the sequencing data using the nf-core RNA-seq pipeline ( https://nf-co.re/rnaseq ). In brief, we performed quality control of the fastq files using FastQC and trimmed the filtered reads with Trim Galore software. The trimmed fastq files resulting from the experiment were aligned to the hg38 human genome using STAR. Salmon was then used to generate a gene-by-sample count matrix for downstream analysis. PCA was performed on read counts that were processed using the variance-stabilizing transformation, and plots were generated from the top 1,000 variable genes across samples. To correct for batch effects by donor, the removeBatchEffect function in the limma package was used. Differential analysis of gene expression was performed using the DESeq2 v.3.16 package, with an absolute log 2 -transformed fold change ≥0.5 and false discovery rate (FDR) < 0.05. To create a heat map, differential genes were aggregated, and expressions were standardized with z -scores across samples. The k -means clustering algorithm with Pearson correlation as the distance metric was used to cluster the genes. Pathway analysis of the differential genes and grouped genes in the heat map was performed using QIAGEN Ingenuity Pathway Analysis 2022 Winter Release and clusterProfiler v.4.6.2. Cell-type enrichment was performed through the single-sample extension of gene set enrichment analysis (ssGSEA) in the GSVA v.1.46.0 R package using signature genes from previous studies 8 , 55 using R v.4.1.0.

Single-cell RNA-seq library preparation and sequencing

To generate single-cell RNA-seq libraries of tumour-infiltrating CAR T cells, Her2 + tumours were collected from five mice per condition, and human CD45 + cells were isolated by NGFR selection as described above (see ‘Cell selection’). Tumour-infiltrating CAR T cells were further purified by sorting human CD3 + TILs from each isolate using a Cytek Aurora Cell Sorter. A total of 20,000 CAR TILs were sorted from each tumour and pooled across five mice per group. Cells were barcoded and sequencing libraries were generated using the 10X Chromium Next GEM Single Cell 3’ v.3.1 kit (10X Genomics) according to the manufacturer’s instructions. Libraries were sequenced at the CHOP High Throughput Sequencing Core on an Illumina NovaSeq 6000 with an average read depth of 50,000 reads per cell.

Single-cell RNA-seq processing and analysis

FASTQ files were generated and aligned to the genome with Cell Ranger v.7.1.0, using a custom GRCh38 reference genome containing the Her2.BBζ CAR sequence. Low-quality cells with fewer than 300 or more than 7,500 genes or more than 10% mitochondrial reads were removed using Seurat v.4.3.0 (ref. 57 ) in R. Doublets were identified using DoubletFinder v.2.0.3 and removed. Filtered samples were normalized using SCTransform before integration. The integrated dataset was scaled, and UMAP dimensionality reduction was performed using the top 30 principal components. Unsupervised Louvain clustering was performed on a shared nearest neighbour graph at a final resolution of 0.6. FindAllMarkers (Seurat) was used to identify DEGs in each cluster, and GO analyses were performed for each cluster using ClusterProfiler v.4.6.2. DEGs and GO processes were used to manually annotate each cluster, and contaminating CD3 − tumour cells were removed. Differential gene analyses between samples were performed using FindMarkers (Seurat) using the Wilcoxon rank-sum test with Bonferroni correction. Gene set scores for T eff , T RM and T reg cell subtypes were calculated with AddModuleScore (Seurat), using curated gene lists from a previous study 58 (Extended Data Fig. 9g–i ). AddModuleScore was also used to calculate a per-cell FOXO1 transcriptional activity score, using the top 100 upregulated genes in CD8 + HA.28ζ CAR T cells overexpressing FOXO1 versus tNGFR (Fig. 2 ). Gene set scores for T eff , T RM and FOXO1 signatures were generated for pan CD3 + T cells (Fig. 4i ; individual genes are shown in Extended Data Fig. 9g–i ). The T reg gene set score was computed for the CD4 + subset of cells expressing ≥1 CD4 mRNA counts and no detectable CD8A counts (Extended Data Fig. 9f ).

Bulk ATAC-seq processing

CD8 + tNGFR + CAR T cells were isolated using the EasySep Human CD8+ T Cell Isolation Kit. A total of 150,000 CD8 + T cells were slow-frozen in BamBanker (Bulldog Bio) cell preservation medium. Approximately 100,000 CAR T cells were washed in ice-cold PBS and subjected to nuclei isolation using the following lysis buffer: 10 mM Tris-HCl pH 7.5, 10 mM NaCl, 3 mM MgCl 2 , 0.1% Tween-20, 0.1% NP40, 0.01% Digitonin and 1% BSA. After washing the cells, 50 μl lysis buffer was added to each sample and cells were resuspended by pipetting. Nuclear pellets were centrifuged and resuspended in the transposase reaction containing 10.5 μl H 2 O, 12.5 μl 2× TD buffer and 2 μl Tn5 transposase in a total of 25 μl. The reaction was incubated for 30 min at 37 °C. The reaction was stopped by the addition of 75 μl TE buffer and 500 μl PB buffer (QIAGEN), followed by column purification per the manufacturer’s recommendation (QIAGEN, Minelute Kit). DNA was eluted from the columns in 22 μl H 2 O. PCR reactions were set up as follows: 21 μl DNA, 25 μl Phusion master mix (NEB) and 2 μl of each barcoded PCR primer (ApexBio, K1058). Fifteen PCR cycles were run for each sample. Reactions were cleaned up with AMPure XP beads according to the recommendations of the manufacturer. Libraries were quantified with a Qubit fluorometer and fragment analysis was performed with Bioanalyzer. Libraries were sequenced on a NovaSeq 6000 sequencer.

Bulk ATAC-seq analysis

ATAC-seq libraries were processed using the pepatac pipeline ( http://pepatac.databio.org/ ) with default options. In brief, fastq files were trimmed to remove adapter sequences, and then pre-aligned to the mitochondrial genome to exclude mitochondrial reads. To ensure the accuracy of downstream analysis, multimapping reads aligning to repetitive regions of the genome were filtered from the dataset. Bowtie2 was then used to align the reads to the hg38 genome. SAMtools was used to identify uniquely aligned reads, and Picard was used to remove duplicate reads. The resulting deduplicated and aligned BAM file was used for downstream analysis. Peaks in individual samples were identified using MACS2 and compiled into a non-overlapping 500-bp consensus peak set. In brief, the peaks were resized to 500 bp width and ranked by significance. The peaks that overlapped with the same region were selected by ranks and the most significant peak was retained. The peak-sample count matrix was generated using ChrAccR with the default parameters of the run_atac function. Signal tracks for individual samples were generated within the pepatac pipeline. These tracks were then merged by group using WiggleTools to produce a comprehensive view of the data across all samples.

On the basis of our analysis of the peak-sample count matrix, the DESeq2 v.3.16 package was used to identify differential peaks across different conditions, with a threshold of an absolute log 2 -transformed fold change greater than 0.5 and P value less than 0.05. Adjusted P values were not used owing to donor variability. To generate PCA plots, we first extracted a variance-stabilized count matrix using the vst function in DESeq2. Next, we corrected for batch effects by donor using the removeBatchEffect function in the limma library. Finally, we generated PCA plots using the corrected matrix with the plotPCA function using the top 2,000 most variable peaks. We aggregated differential peaks across conditions, standardized the peak signals using z -scores across samples and performed k -means clustering to generate a chromatin accessibility heat map. Motif enrichments of differential peaks and grouped peaks were searched with HOMER and findMotifsGenome.pl with default parameters. The enrichment of cell-type-specific regulatory elements were performed with the gchromVAR package. In brief, this method weights chromatin features by log 2 -transformed fold changes of cell-type-specific regulatory elements from a previous report 9 and computes the enrichment for each cell type versus an empirical background matched for GC content and feature intensity.

Identification and analysis of the FOXO1 regulon

The FOXO1 regulon gene set was generated by intersecting downregulated differential genes (log 2 -transformed fold change < −0.25, FDR < 0.05) in FOXO1 KO cells and upregulated differential genes (log 2 -transformed fold change > 0.5, FDR < 0.05) in FOXO1 OE cells (Supplementary Table 1 ). Regulon enrichment scores were calculated using ssGSEA in the GSVA R package on a previous RNA expression dataset 2 .

For regulon analyses of single-cell ATAC-seq data, the processed Signac data objects of CAR T products profiled by single-cell ATAC-seq were obtained from a previous study 5 . To account for sample-to-sample variability, the mean fragments in peaks per cell were downsampled for consistency between donors. Furthermore, donors PT48 and PT51 were excluded on the basis of low data quality after examination of quality control statistics, including per-library transcription start site enrichment. Using the epigenetic signature for FOXO1 and TCF1 overexpression (Fig. 2 ), we computed the per-cell epigenetic signature per factor using the chromVAR workflow as previously described for related T cell signatures derived from bulk experiments. To test for differences in responder/non-responder associations with this signature, we performed an ordinary least squares regression with the per-cell z -score against the donor’s BCA status at 6 months, adjusting for individual patient ID. Statistical significance was based on the Wald test statistic of the coefficient for the responder term in the two regressions for each factor.

For regulon analyses of the CLL CD19 CAR T cell clinical dataset, the gene-expression data table for activated CD19 CAR T cell products from patients with CLL was obtained from a previous report 2 . The enrichment of the FOXO1 signature was analysed using ssGSEA as previously described and performed using the R package GSVA v.1.46.0. To compare the ssGSEA enrichment scores between responders and non-responders, a Mann–Whitney test was conducted. To statistically determine optimal stratification points for survival analysis, we compared candidate stratification points on the basis of hazard ratio and P value as previously described. The survival analysis was conducted with a log-rank (Mantel–Cox) test using GraphPad Prism v.9.5.0.

Statistical analyses

Unless otherwise stated, statistical analyses for significant differences between groups were conducted using one- or two-way analysis of variance (ANOVA) with Bonferroni, Tukey’s or Dunnett’s multiple comparisons test, or with a Student’s or Welch’s t -test using GraphPad Prism v.9.4.1. In experiments in which same-donor samples were compared across two conditions, we performed a paired Student’s t -test. Survival curves were compared using the log-rank Mantel–Cox test. Statistical methods were not used to predetermine sample sizes.

Reporting summary

Further information on research design is available in the  Nature Portfolio Reporting Summary linked to this article.

Data availability

Transcription factor constructs will be made available through material transfer agreements when possible. The bulk RNA-seq, ATAC-seq and single-cell RNA-seq datasets were aligned to human genome hg38; they have been deposited in the NCBI Gene Expression Omnibus and are accessible through the accession number GSE255416 .  Source data are provided with this paper.

Code availability

All code associated with this paper have been deposited to the Weber Lab GitHub repository ( https://github.com/Weber-Lab-CHOP/FOXO1_2024 ) 59 .

Weber, E. W., Maus, M. V. & Mackall, C. L. The emerging landscape of immune cell therapies. Cell 181 , 46–62 (2020).

Article   CAS   PubMed   PubMed Central   Google Scholar  

Fraietta, J. A. et al. Determinants of response and resistance to CD19 chimeric antigen receptor (CAR) T cell therapy of chronic lymphocytic leukemia. Nat. Med. 24 , 563–571 (2018).

Deng, Q. et al. Characteristics of anti-CD19 CAR T cell infusion products associated with efficacy and toxicity in patients with large B cell lymphomas. Nat. Med. 26 , 1878–1887 (2020).

Bai, Z. et al. Single-cell antigen-specific landscape of CAR T infusion product identifies determinants of CD19-positive relapse in patients with ALL. Sci. Adv. 8 , eabj2820 (2022).

Chen, G. M. et al. Integrative bulk and single-cell profiling of premanufacture T-cell populations reveals factors mediating long-term persistence of CAR T-cell therapy. Cancer Discov. 11 , 2186–2199 (2021).

Kirouac, D. C. et al. Deconvolution of clinical variance in CAR-T cell pharmacology and response. Nat. Biotechnol. 41 , 1606–1617 (2023).

Weber, E. W. et al. Transient rest restores functionality in exhausted CAR-T cells through epigenetic remodeling. Science 372 , eaba1786 (2021).

Krishna, S. et al. Stem-like CD8 T cells mediate response of adoptive cell immunotherapy against human cancer. Science 370 , 1328–1334 (2020).

Article   ADS   CAS   PubMed   PubMed Central   Google Scholar  

Satpathy, A. T. et al. Massively parallel single-cell chromatin landscapes of human immune cell development and intratumoral T cell exhaustion. Nat. Biotechnol. 37 , 925–936 (2019).

Sade-Feldman, M. et al. Defining T Cell states associated with response to checkpoint immunotherapy in melanoma. Cell 175 , 998–1013 (2018).

Fabre, S. et al. FOXO1 regulates L-selectin and a network of human T cell homing molecules downstream of phosphatidylinositol 3-kinase. J. Immunol. 181 , 2980–2989 (2008).

Article   CAS   PubMed   Google Scholar  

Kim, M. V., Ouyang, W., Liao, W., Zhang, M. Q. & Li, M. O. The transcription factor Foxo1 controls central-memory CD8 + T cell responses to infection. Immunity 39 , 286–297 (2013).

Hess Michelini, R., Doedens, A. L., Goldrath, A. W. & Hedrick, S. M. Differentiation of CD8 memory T cells depends on Foxo1. J. Exp. Med. 210 , 1189–1200 (2013).

Article   PubMed   Google Scholar  

Utzschneider, D. T. et al. Active maintenance of T cell memory in acute and chronic viral infection depends on continuous expression of FOXO1. Cell Rep. 22 , 3454–3467 (2018).

Klebanoff, C. A. et al. Inhibition of AKT signaling uncouples T cell differentiation from expansion for receptor-engineered adoptive immunotherapy. JCI Insight 2 , e95103 (2017).

Article   PubMed   PubMed Central   Google Scholar  

Crompton, J. G. et al. Akt inhibition enhances expansion of potent tumor-specific lymphocytes with memory cell characteristics. Cancer Res. 75 , 296–305 (2015).

Mehra, V. et al. AKT inhibition generates potent polyfunctional clinical grade AUTO1 CAR T-cells, enhancing function and survival. J. Immunother. Cancer 11 , e007002 (2023).

Nagashima, T. et al. Discovery of novel forkhead box O1 inhibitors for treating type 2 diabetes: improvement of fasting glycemia in diabetic db / db mice. Mol. Pharmacol. 78 , 961–970 (2010).

Staron, M. M. et al. The transcription factor FoxO1 sustains expression of the inhibitory receptor PD-1 and survival of antiviral CD8 + T cells during chronic infection. Immunity 41 , 802–814 (2014).

Delpoux, A. et al. Continuous activity of Foxo1 is required to prevent anergy and maintain the memory state of CD8 + T cells. J. Exp. Med. 215 , 575–594 (2018).

Delpoux, A., Lai, C.-Y., Hedrick, S. M. & Doedens, A. L. FOXO1 opposition of CD8 + T cell effector programming confers early memory properties and phenotypic diversity. Proc. Natl Acad. Sci. USA 114 , E8865–E8874 (2017).

Delpoux, A. et al. FOXO1 constrains activation and regulates senescence in CD8 T cells. Cell Rep. 34 , 108674 (2021).

Lynn, R. C. et al. c-Jun overexpression in CAR T cells induces exhaustion resistance. Nature 576 , 293–300 (2019).

Weber, E. W. et al. Pharmacologic control of CAR-T cell function using dasatinib. Blood Adv. 3 , 711–717 (2019).

Im, S. J. et al. Defining CD8 + T cells that provide the proliferative burst after PD-1 therapy. Nature 537 , 417–421 (2016).

Siddiqui, I. et al. Intratumoral Tcf1 + PD-1 + CD8 + T Cells with stem-like properties promote tumor control in response to vaccination and checkpoint blockade immunotherapy. Immunity 50 , 195–211 (2019).

Shan, Q. et al. Ectopic Tcf1 expression instills a stem-like program in exhausted CD8 + T cells to enhance viral and tumor immunity. Cell. Mol. Immunol. 18 , 1262–1277 (2021).

Wu, T. et al. The TCF1–Bcl6 axis counteracts type I interferon to repress exhaustion and maintain T cell stemness. Sci. Immunol. 1 , eaai8593 (2016).

Tsui, C. et al. MYB orchestrates T cell exhaustion and response to checkpoint inhibition. Nature 609 , 354–360 (2022).

Wang, Y. et al. The transcription factor TCF1 preserves the effector function of exhausted CD8 T cells during chronic viral infection. Front. Immunol. 10 , 169 (2019).

Giuffrida, L. et al. IL-15 preconditioning augments CAR T cell responses to checkpoint blockade for improved treatment of solid tumors. Mol. Ther. 28 , 2379–2393 (2020).

Hudson, W. H. et al. Proliferating transitory T Cells with an effector-like transcriptional signature emerge from PD-1 + stem-like CD8 + T cells during chronic infection. Immunity 51 , 1043–1058 (2019).

Gattinoni, L. et al. Wnt signaling arrests effector T cell differentiation and generates CD8 + memory stem cells. Nat. Med. 15 , 808–813 (2009).

Kerdiles, Y. M. et al. Foxo transcription factors control regulatory T cell development and function. Immunity 33 , 890–904 (2010).

Ouyang, W. et al. Novel Foxo1-dependent transcriptional programs control T reg cell function. Nature 491 , 554–559 (2012).

Shan, Q. et al. Tcf1 and Lef1 provide constant supervision to mature CD8 + T cell identity and function by organizing genomic architecture. Nat. Commun. 12 , 5863 (2021).

Hatta, M. & Cirillo, L. A. Chromatin opening and stable perturbation of core histone:DNA contacts by FoxO1. J. Biol. Chem. 282 , 35583–35593 (2007).

Miller, B. C. et al. Subsets of exhausted CD8 + T cells differentially mediate tumor control and respond to checkpoint blockade. Nat. Immunol. 20 , 326–336 (2019).

Chen, Z. et al. TCF-1-centered transcriptional network drives an effector versus exhausted CD8 T cell-fate decision. Immunity 51 , 840–855 (2019).

Farber, D. L., Yudanin, N. A. & Restifo, N. P. Human memory T cells: generation, compartmentalization and homeostasis. Nat. Rev. Immunol. 14 , 24–35 (2014).

Matsuzaki, H. et al. Acetylation of Foxo1 alters its DNA-binding ability and sensitivity to phosphorylation. Proc. Natl Acad. Sci. USA 102 , 11278–11283 (2005).

Miyara, M. et al. Functional delineation and differentiation dynamics of human CD4 + T cells expressing the FoxP3 transcription factor. Immunity 30 , 899–911 (2009).

Bonacci, B. et al. Requirements for growth and IL-10 expression of highly purified human T regulatory cells. J. Clin. Immunol. 32 , 1118–1128 (2012).

Johnson, J. L. et al. Lineage-determining transcription factor TCF-1 initiates the epigenetic identity of T cells. Immunity 48 , 243–257 (2018).

Wang, W. et al. TCF-1 promotes chromatin interactions across topologically associating domains in T cell progenitors. Nat. Immunol. 23 , 1052–1062 (2022).

Gray, S. M., Amezquita, R. A., Guan, T., Kleinstein, S. H. & Kaech, S. M. Polycomb repressive complex 2-mediated chromatin repression guides effector CD8 + T cell terminal differentiation and loss of multipotency. Immunity 46 , 596–608 (2017).

Oh, H.-M. et al. STAT3 protein promotes T-cell survival and inhibits interleukin-2 production through up-regulation of Class O Forkhead transcription factors. J. Biol. Chem. 286 , 30888–30897 (2011).

Rao, R. R., Li, Q., Gubbels Bupp, M. R. & Shrikant, P. A. Transcription factor Foxo1 represses T-bet-mediated effector functions and promotes memory CD8 + T cell differentiation. Immunity 36 , 374–387 (2012).

Tejera, M. M., Kim, E. H., Sullivan, J. A., Plisch, E. H. & Suresh, M. FoxO1 controls effector-to-memory transition and maintenance of functional CD8 T cell memory. J. Immunol. 191 , 187–199 (2013).

Ouyang, W., Beckett, O., Flavell, R. A. & Li, M. O. An essential role of the Forkhead-box transcription factor Foxo1 in control of T cell homeostasis and tolerance. Immunity 30 , 358–371 (2009).

Zhang, L. et al. Mammalian target of rapamycin complex 2 controls CD8 T cell memory differentiation in a Foxo1-dependent manner. Cell Rep. 14 , 1206–1217 (2016).

Smole, A. et al. Expression of inducible factors reprograms CAR-T cells for enhanced function and safety. Cancer Cell 40 , 1470–1487 (2022).

Moffett, H. F. et al. Hit-and-run programming of therapeutic cytoreagents using mRNA nanocarriers. Nat. Commun. 8 , 389 (2017).

Roth, T. L. et al. Pooled knockin targeting for genome engineering of cellular immunotherapies. Cell 181 , 728–744 (2020).

Andreatta, M. et al. Interpretation of T cell states from single-cell transcriptomics data using reference atlases. Nat. Commun. 12 , 2965 (2021).

Jung, I.-Y. et al. BLIMP1 and NR4A3 transcription factors reciprocally regulate antitumor CAR T cell stemness and exhaustion. Sci. Transl. Med. 14 , eabn7336 (2022).

Hao, Y. et al. Integrated analysis of multimodal single-cell data. Cell 184 , 3573–3587 (2021).

Bukhari, S. et al. Single-cell RNA sequencing reveals distinct T cell populations in immune-related adverse events of checkpoint inhibitors. Cell Rep. Med. 4 , 100868 (2023).

Weber, E .W. Weber Lab. GitHub https://github.com/Weber-Lab-CHOP/FOXO1_2024 (2024).

Jung, I.-Y. et al. Type I interferon signaling via the EGR2 transcriptional regulator potentiates CAR T cell-intrinsic dysfunction. Cancer Discov. 13 , 1636–1655 (2023).

Download references

Acknowledgements

We thank W. Yu for guidance with single-cell ATAC-seq data analysis; R. Majzner and F. Staback Rodriguez for thoughtful discussions; the National Cancer Institute at Frederick for providing the 1A7 anti-14g2a idiotype antibody; B. Jena and L. J. N. Cooper for providing the monoclonal anti-FMC63 idiotype antibody; and the High Throughput Sequencing Core at CHOP for help with sequencing. We also acknowledge the Flow Cytometry Core at CHOP and the Stanford Institute for Stem Cell Biology and Regenerative Medicine FACS core for equipment and technical support. Schematics were created with BioRender.com. This work was supported by the National Cancer Institute Immunotherapy Discover and Development (1U01CA232361-A1 to S.A.G. and E.W.W.; K08CA23188-01 and U01CA260852 to A.T.S.; and U54CA232568-01 to C.L.M.); the National Human Genome Research Institute (K99 HGHG012579 to C.A.L.); the Parker Institute for Cancer Immunotherapy (C.A.L., A.T.S., C.L.M. and E.W.W); the V Foundation for Cancer Research (E.W.W.); a Society for Immunotherapy of Cancer Rosenberg Scholar Award (E.W.W.); Stand Up 2 Cancer (St. Baldrick’s) (NCI SU2CAACR-DT1113 to C.L.M.); the Virginia and D.K. Ludwig Fund for Cancer Research (C.L.M.); and NCI U2C CA233285 (K.T.). C.L.M., A.T.S., C.A.L. and E.W.W. are members of the Parker Institute for Cancer Immunotherapy, which supports cancer immunotherapy research at Stanford University and the University of Pennsylvania. Stand Up 2 Cancer is a program of the Entertainment Industry Foundation administered by the American Association for Cancer Research.

Author information

Bence Daniel

Present address: Genentech, South San Francisco, CA, USA

These authors contributed equally: Alexander E. Doan, Katherine P. Mueller, Andy Y. Chen

These authors jointly supervised this work: Crystal L. Mackall, Evan W. Weber

Authors and Affiliations

Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA, USA

Alexander E. Doan, John Lattin, Peng Xu, Dorota Klysz, Malek Bashti, Patrick J. Quinn, Elena Sotillo & Crystal L. Mackall

Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

Katherine P. Mueller, Geoffrey T. Rouin, Yingshi Chen, Martina Markovska, Brett Mozarsky, Jose Arias-Umana, Robert Hapke, Alice Wang, Gabrielle Zuern, Faith Ryu, Junior Hall, Kai Tan, Stephan A. Grupp, Susan E. McClory & Evan W. Weber

Center for Cellular and Molecular Therapeutics, Children’s Hospital of Philadelphia, Philadelphia, PA, USA

Katherine P. Mueller, Geoffrey T. Rouin, Yingshi Chen, Martina Markovska, Brett Mozarsky, Jose Arias-Umana, Robert Hapke, Gabrielle Zuern, Faith Ryu & Evan W. Weber

Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

Katherine P. Mueller, Geoffrey T. Rouin, Yingshi Chen, Martina Markovska, Brett Mozarsky, Jose Arias-Umana, Robert Hapke, Gabrielle Zuern, Gregory M. Chen, Faith Ryu, Meghan Logun, Joseph A. Fraietta & Evan W. Weber

Department of Pathology, Stanford University, Stanford, CA, USA

Andy Y. Chen, Bence Daniel, Katalin Sandor, Wenxi Zhang, Caleb A. Lareau & Ansuman T. Satpathy

Department of Bioengineering, Stanford University, Stanford, CA, USA

Andy Y. Chen

Gladstone–UCSF Institute of Genomic Immunology, San Francisco, CA, USA

Center for Personal Dynamic Regulomes, Stanford University, Stanford, CA, USA

Department of Genetics, Stanford University, Stanford, CA, USA

Bence Daniel & Zhuang Miao

Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

In-Young Jung & Joseph A. Fraietta

Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

Gregory M. Chen & Joseph A. Fraietta

Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

Meghan Logun

Center for Childhood Cancer Research, Children’s Hospital of Philadelphia, Philadelphia, PA, USA

Junior Hall, Kai Tan, Stephan A. Grupp, Susan E. McClory & Evan W. Weber

Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

Junior Hall, Kai Tan, Stephan A. Grupp, Joseph A. Fraietta & Evan W. Weber

Parker Institute for Cancer Immunotherapy, San Francisco, CA, USA

Caleb A. Lareau, Ansuman T. Satpathy, Crystal L. Mackall & Evan W. Weber

Department of Pediatrics, Stanford University, Stanford, CA, USA

Crystal L. Mackall

Department of Medicine, Stanford University, Stanford, CA, USA

You can also search for this author in PubMed   Google Scholar

Contributions

E.W.W. and C.L.M. conceived the study, secured funding and supervised the project. A.E.D., K.P.M., G.T.R., B.D., J.L., Y.C., B.M., M.M., J.A.-U., R.H., A.W., P.X., D.K., G.Z., M.B., P.J.Q., Z.M., K.S., W.Z., F.R., M.L., J.H., S.E.M. and E.W.W. designed and performed wet-lab experiments. K.P.M., A.Y.C., B.D., A.W., G.M.C., and C.A.L. performed RNA-seq and ATAC-seq computational analyses. I.-Y.J. and J.A.F. performed analyses on and interpreted clinical data. K.T., S.A.G., J.A.F., E.S. and A.T.S. supervised experiments and analyses. A.E.D., K.P.M., E.S., C.L.M. and E.W.W. wrote the manuscript. All authors discussed the results and edited the manuscript.

Corresponding authors

Correspondence to Crystal L. Mackall or Evan W. Weber .

Ethics declarations

Competing interests.

C.A.L. is a consultant to Cartography Biosciences. S.A.G. receives research funding from Novartis, Kite, Vertex and Servier; consults for Novartis, Roche, GSK, Humanigen, CBMG, Eureka, Janssen/JNJ and Jazz Pharmaceuticals; and has advised for Novartis, Adaptimmune, TCR2, Cellctis, Juno, Vertex, Allogene, Jazz Pharmaceuticals and Cabaletta. J.A.F. receives research funding from Tceleron (formerly Tmunity Therapeutics) and Danaher Corporation; consults for Retro Biosciences; and is a member of the scientific advisory boards of Cartography Biosciences and Shennon Biotechnologies. A.T.S. is a founder of Immunai and Cartography Biosciences and receives research funding from Allogene Therapeutics and Merck Research Laboratories. C.L.M. is a co-founder of and holds equity in Link Cell Therapies, Cargo Therapeutics (formerly Syncopation Life Sciences) and Lyell Immunopharma; holds equity and consults for Mammoth and Ensoma; consults for Immatics and Nektar; and receives research funding from Tune Therapeutics. E.W.W. holds equity in Lyell Immunopharma and consults for Umoja Immunopharma. The remaining authors declare no competing interests.

Peer review

Peer review information.

Nature thanks the anonymous reviewer(s) for their contribution to the peer review of this work. Peer reviewer reports are available.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data figures and tables

Extended data fig. 1 pharmacological inhibition of foxo1 impairs expansion, formation of a memory phenotype and antitumour function in cd19.28ζ and cd19.bbζ car t cells..

CAR T cells were treated with DMSO or 10 nM or 100 nM of the small molecule AS1842856 (FOXO1 i ) starting on day 4 post-activation and treated every 2–3 days thereafter. a , Schematic of FOXO1 i experimental model. b , CD19.28ζ (left) or CD19.BBζ (right) CAR T cell expansion ( n  = 2 donors). c , Percent CD8 + in CD19.28ζ (circles) and CD19.BBζ (squares) cells ( n  = 2 donors for each CAR). d , Apoptosis in CD19.BBζ CAR T cells at day 15 post-activation. Contour plots show 1 representative donor and bar graphs show mean±s.e.m. of n  = 3 donors. e , Expression of memory- and exhaustion-associated markers on CD19.28ζ and CD19.BBζ cells. Histograms show 1 representative donor ( n  = 2 donors). f , Cytokine secretion from CD19.28ζ and CD19.BBζ cells in response to Nalm6 cells. Graphs show mean±s.d. of triplicate wells from 1 representative donor ( n  = 2 donors). g , Cytotoxicity of CD19.BBζ cells against Nalm6 cells at a 1:1 E:T ratio. Data is normalized to t  = 0 and show mean±s.d. of triplicate wells from 1 representative donor ( n  = 2 donors). Statistics are shown for t  = 60 h. Statistical comparisons were performed using paired two-tailed Student’s t -test ( c ), two-way ANOVA with Šídák’s test ( d ) and one-way ( f ) or two-way ( g ) ANOVA with Dunnett’s test. E:T ratio, effector:target cell ratio. NS, not significant.

Extended Data Fig. 2 CRISPR knockout of FOXO1 attenuates memory formation and promotes exhaustion in CD19.BBζ and HA.28ζ CAR T cells.

a – i , CRISPR–Cas9 gene editing of AAVS1 (AAVS1) or FOXO1 (FOXO1 KO ) in CD19.BBζ CAR T cells. a , Schematic depicting generation of FOXO1 KO CAR T cells and downstream assays. b , Day 14 FOXO1 KO expansion normalized to AAVS1. Data show mean±s.e.m. of n  = 3 donors. c – f , Flow cytometric analysis of memory- and exhaustion-associated markers on CD8 + ( c , e ) and CD4 + ( d , f ) CD19.BBζ cells. Histograms and contour plots show a representative donor and bar graphs show mean±s.e.m. of n  = 3-6 donors. CD62L, IL-7Rα, TCF1, and CD39 histograms in c also appear in Fig. 1c . g , MFI of CD62L in FOXO1 + and FOXO1 − gated subpopulations of CD19.BBζ cells. h , Schematic showing CD62L lo / FOXO1 KO cell negative selection strategy for RNA-seq experiments. i , GO term analyses showing curated lists of up- and downregulated processes in FOXO1 KO compared to AAVS1. Data show Benjamini–Hochberg-adjusted P value ( n  = 3 donors). j , Flow cytometric analysis of memory- and exhaustion-associated markers in day 15 HA.28ζ CAR T cells. Background-subtracted MFI is displayed. k , Cytokine secretion from day 15 HA.28ζ cells in response to Nalm6. Graphs show mean±s.d. of 3 technical replicates from one representative donor ( n  = 2 donors). Statistical comparisons were performed using paired two-tailed Student’s t -test ( b , c , d , g ), two-way ANOVA with Bonferroni’s test ( e , f ), two-tailed Student’s t -test ( k ) and one-sided hypergeometric test ( i ).

Extended Data Fig. 3 FOXO1 overexpression promotes a memory phenotype and mitigates exhaustion in CAR T cells.

a , Schematic depicting engineering of truncated NGFR-only (tNGFR), TCF1/tNGFR- (TCF1 OE ), and FOXO1/tNGFR- (FOXO1 OE ) CAR T cells and magnetic isolation of tNGFR + cells for downstream analyses. b – e , Phenotypic and functional analyses of CD19.BBζ CAR T cells at baseline and during repeat stimulation with Nalm6 cells. b , c , Flow cytometric analysis of CD62L and IL-7Rα ( b ) and TCF1 and LEF1 ( c ) from 1 representative donor ( n  = 4 donors). d , Cytokine secretion from CD19.BBζ cells after 1 or 4 stimulations with Nalm6 cells. Data show mean ± s.d. of 2–3 triplicate wells from 1 representative donor ( n  = 2 donors). e , Flow cytometric analysis of CD62L, IL-7Rα, and CD39 on tNGFR + CD8 + CAR T cells prior to the first stimulation (Stim 0) and 7 days after the third stimulation (Stim 3). Data show mean ± s.e.m. of mean fluorescence intensity normalized to tNGFR levels from n  = 2–3 donors. f – i , CAR T cell exhaustion model 7 , 23 whereby T cells express a high-affinity GD2-targeting CAR (HA.28ζ) that promotes antigen-independent tonic CAR signalling. f , Model schematic. g , Flow cytometric analysis of day 15 CD62L and IL-7Rα. Data show 1 representative donor ( n  = 5 donors). h , Cytotoxicity of day 15 HA.28ζ cells against 143B cells at a 1:8 E:T ratio. Data is normalized to t  = 0 and show mean±s.d. of 3 triplicate wells from 1 representative donor ( n  = 3 donors). Statistics were performed at t  = 96 h. i , j , Seahorse metabolic analyses on day 13 of culture ( n  = 2 donors). i , Ratio of OCR to ECAR of HA.28ζ cells. j , CD19.28ζ cell OCR (left), OCR to ECAR ratio (centre), and spare respiratory capacity (right). OCR line graph shows 1 representative donor. Bar graphs show mean ± s.d. of three representative time points within each donor. Statistical comparisons were performed using one-way ANOVA with Tukey’s test ( h ) or Dunnett’s test ( i , j ). E:T ratio, effector:target cell ratio. OCR, Oxygen consumption rate. ECAR, extracellular acidification rate.

Extended Data Fig. 4 Overexpression of FOXO1 induces a memory-like transcriptional program in CAR T cells.

a – g , Bulk RNA-seq analyses of day 15 tNGFR + CD4 + HA.28ζ CAR T cells overexpressing tNGFR, TCF1 OE , or FOXO1 OE ( n  = 3 donors). a , PCA of CD4 + cells. b , PCA that includes CD4 + samples plotted in a and CD8 + samples plotted in Fig. 2a . c , Venn diagram showing the number of unique and shared DEGs in CD4 + TCF1 OE and FOXO1 OE cells compared to tNGFR cells (Bonferroni-adjusted P  < 0.05 with abs(log 2 FC)>0.5). d , Expression of memory- and exhaustion-associated genes. Centre line represents the mean counts per million. e , Heat map and hierarchical clustering of DEGs. Genes of interest are shown. Scale shows normalized z-scores for each DEG. f , GSVA using published human CD4 + regulatory T cell (T reg ) signatures 42 , 43 . Centre line represents mean score. g , GO term analyses showing curated lists of top up- and downregulated processes in CD4 + FOXO1 OE and TCF1 OE cells versus tNGFR cells. Data show Benjamini–Hochberg-adjusted P . h , QIAGEN IPA of upregulated and downregulated TF pathways in FOXO1 OE cells versus tNGFR cells. Data show adjusted P . i – k , Bulk RNA-seq analyses of day 15 tNGFR + CD8 + CD19.28ζ cells overexpressing tNGFR, TCF1 OE , or FOXO1 OE ( n  = 3 donors). i , PCA analysis. j , Venn diagram showing the number of unique and shared DEGs in TCF1 OE and FOXO1 OE cells compared to tNGFR cells (Bonferroni-adjusted P  < 0.05 with log 2 (fold change) < 0.5). k , Heat map and hierarchical clustering of DEGs. Genes of interest are shown. Scale shows normalized z-scores for each DEG. Statistical comparisons were performed using DESeq2 ( c , d , e , j , k ), repeated-measures one-way ANOVA with Tukey’s test ( f ) and one-sided hypergeometric test ( g ). NS, not significant.

Extended Data Fig. 5 FOXO1 or TCF1 overexpression induces chromatin remodelling in CD19.28ζ and HA.28ζ CAR T cells.

a – e , Bulk ATAC-seq analyses of day 15 tNGFR + CD8 + CAR T cells expressing either CD19.28ζ ( a – d ) or HA.28ζ ( e ) ( n  = 3 donors). a , PCA of CD19.28ζ cells. b , c , Rank-ordered plot of differentially accessible TF-binding motifs ( P  < 0.05 with abs(log 2 FC)>0.5) in FOXO1 OE cells ( b ) and TCF1 OE cells ( c ) versus tNGFR cells. d , e , Heat maps and hierarchical clustering of mean differential motif accessibility of CD19.28ζ ( d ) or HA.28ζ ( e ) cells. Scales show normalized z-scores for each motif. Statistical comparisons were performed using DESeq2 ( b – e ).

Extended Data Fig. 6 Nuclear-restricted FOXO1 promotes a memory-like phenotype but impairs effector function.

a , Schematic showing a mutated variant of FOXO1 that contains three amino acid substitutions (T24A, S256A, and S319A) which restrict nuclear export (FOXO1 3A ). b , Analysis of soluble and chromatin-bound FOXO1 fractions isolated from tNGFR + non-CAR T cells that were activated with Dynabeads for 24 h prior to cell collection. Western blots (left) and bar graph (right) representing the ratio of chromatin-bound to soluble FOXO1 normalized to mock T cells are shown for 1 representative donor ( n  = 2 donors). c , FOXO1 expression in CD19.28ζ and HA.28ζ CAR T cells from 1 representative donor ( n  = 5 donors). d , CD62L and IL-7Rα expression in CD19.28ζ and HA.28ζ CAR T cells from 1 representative donor ( n  = 3 donors). e , TCF1 and LEF1 expression in CD19.28ζ CAR T cells from 1 representative donor ( n  = 3 donors). f , g , RNA-seq on HA.28ζ CAR T cells. tNGFR and FOXO1 OE samples are also represented in Fig. 2 and Extended Data Fig. 4 . f , PCA. g , GO term analyses showing curated lists of top up- and downregulated processes in FOXO1 3A vs FOXO1 OE . Data show Benjamini–Hochberg-adjusted P value. h , Cytotoxicity of HA.28ζ cells against Nalm6 at a 1:1 E:T ratio. Data are normalized to t  = 0 and show mean±s.d. from 1 representative donor ( n  = 3 donors). Statistics were performed at t  = 96 h. i , Cytokine secretion from day 15 HA.28ζ CAR T cells in response to 143B cells. Plots show mean±s.d. of 3 wells from 1 representative donor ( n  = 3 donors). Statistical comparisons were performed using one-sided hypergeometric test ( g ), one-way ANOVA with Tukey’s test ( h ) or Dunnett’s test ( i ). E:T ratio, effector:target cell ratio.

Extended Data Fig. 7 FOXO1 OE CAR T cells show enhanced antitumour activity in leukaemia xenograft models .

a , b , A curative dose of 2×10 6 tNGFR + CD19.BBζ CAR T cells overexpressing tNGFR, TCF1 OE , FOXO1 OE , or FOXO1 3A were infused into Nalm6 leukaemia-bearing mice 7 days post-engraftment ( n  = 2 donors tested in 2 independent experiments) a , Experimental schematic (left) and tumour bioluminescence of multiple time points (right) from 1 representative donor ( n  = 3-5 mice per group). b , Tumour bioluminescence from day 42-45. Data show mean±s.e.m. from 2 donors tested in 2 independent experiments ( n  = 3-10 mice per group; n  = 1 donor for FOXO1 3A ). c – g , A curative dose of 1×10 6 tNGFR + CD19.28ζ cells were infused into Nalm6-bearing mice 7 days post-engraftment. Mice were rechallenged with 10×10 6 CD19 + or CD19 − Nalm6 on day 21 post-CAR T cell infusion ( n  = 2 donors tested in 2 independent experiments). c , Tumour bioluminescence over time. Data show mean±s.e.m. of n  = 3–7 mice per group from 1 representative donor. d , CD19.28ζ and tNGFR expression on circulating CD45 + CAR T cells on day 21. Contour plots show 1 representative mouse from each condition from 1 representative donor. e , Quantification of circulating CD45 + CAR T cells on days 7, 21, and 28. f , CD4 + and CD8 + CAR T cells on day 7 (data derived from e ). g , Percent CD8 + CAR T cells. Graphs in e – g show mean±s.e.m. of n  = 3–7 mice per group from 1 representative donor. Statistical comparisons were performed using nonparametric two-tailed Mann–Whitney test ( b ) and two-way ( c ) and one-way ANOVA with Dunnett’s test ( e , f ) and mixed-effects model with Dunnett’s test ( g ). NS, not significant.

Extended Data Fig. 8 FOXO1 OE reprogramming and enhanced antitumour activity are dependent on DNA binding.

a , Schematic depicting construct design and amino acid substitutions (K245A and K248A) to generate human FOXO1 DBD (top) and western blots of indicated proteins in soluble and chromatin-bound fractions isolated from day 8 tNGFR + CD19.28ζ CAR T cells (bottom). Densitometry analyses are displayed below the blots. 1 representative donor from n  = 2 donors. b , FOXO1 expression in CD19.28ζ CAR T cells from one representative donor ( n  = 5 donors). c , Bulk RNA-seq analyses of day 15 tNGFR + CD8 + HA.28ζ CAR T cells show unique and shared DEGs in FOXO1 DBD  and FOXO1 OE compared with tNGFR (Bonferroni-adjusted P  < 0.05 with abs(log 2 FC)>0.5). FOXO1 OE samples are also represented in Fig. 2 and Extended Data Figs. 4 and 6 . d , Bulk ATAC-seq of day 15 tNGFR + CD8 + HA.28ζ CAR T cells. Rank-ordered plot of differentially accessible TF-binding motifs in FOXO1 OE cells versus FOXO1 DBD cells ( P  < 0.05 with abs(log 2 FC)>0.5). FOXO1 OE samples are also represented in Fig. 2 and Extended Data Fig. 5 . e , Schematic of stress test model (left) whereby Nalm6-engrafted mice were treated with mock T cells or FOXO1 OE or FOXO1 DBD CD19.28ζ CAR T cells. Survival curve shows pooled data from 2 donors tested in 2 independent experiments ( n  = 10 mice per group, FOXO1 OE data from 1 donor are also represented in Fig. 3a ). f , TCF7 knockout efficiency for bulk RNA-seq data corresponding to Fig. 3f,g . Data show the mean of n  = 3 donors with 2 technical replicates per donor. Statistical comparisons were performed using DESeq2 ( c , d ) and log-rank Mantel–Cox test ( e ).

Extended Data Fig. 9 FOXO1 OE CAR T cells exhibit improved persistence and effector- and tissue-residence-associated transcriptomic signatures in a solid-tumour xenograft model.

5×10 6 Her2.BBζ CAR T cells were infused into 143B-bearing mice 3 days post-engraftment. Tumours and spleens were collected on day 29 post-engraftment for phenotypic, functional, and sequencing-based assays. a , Total splenic CAR T cells. b , Total CD4 + (left) and CD8 + (right) splenic CAR T cells. c , Ratio of CD8 + to CD4 + tumour-infiltrating CAR T cells from donor 1 ( n  = 3 mice per group). Donor 2 is shown in Fig. 4d . d , Ratio of CD8 + to CD4 + CAR T cells from spleens. Data in a – d show mean±s.d. of n  = 13 mice per group from 2 donors tested in 2 independent experiments unless otherwise stated. e – i , Single-cell RNA-seq on day 29 tumour-infiltrating FOXO1 OE or tNGFR cells. Cells were sorted and pooled from n  = 5 mice per group from 1 donor. e , Top enriched GO terms in Cluster 1, which was biased towards FOXO1 OE cells. Gene ratio and Benjamini–Hochberg-adjusted P value are shown. f , T reg transcriptional signature 58 score. g , T eff signature genes corresponding to T eff scores in Fig. 4i . h , T RM signature genes corresponding to T RM scores in Fig. 4i . i , T reg signature genes corresponding to scores in f . Statistical comparisons were performed using two-tailed Student’s t -test ( a-d ), one-sided hypergeometric test ( e ) and two-sided Wilcoxon rank-sum test ( f ).

Extended Data Fig. 10 Endogenous TCF7 transcript and FOXO1 regulon, but not TCF OE transcriptional or epigenetic signatures, predict CAR T cell and TIL responses in patients.

a , ssGSEA on RNA-seq from CAR-stimulated CTL019 cells 2 (complete responder, CR, partial responder with transformed disease, PR TD , n  = 3; partial responder, PR, n  = 5; non-responder, NR, n  = 21). Enrichment score stratification points for patient survival analyses were determined using previously published methods 60 . a , TCF7 expression is shown for patient outcomes (left) and overall survival (right). b – d , P values (top) and hazard ratios (bottom) of different stratification points in relation to overall survival (OS) of TCF7 expression ( b ), FOXO1 expression ( c ) and FOXO1 regulon ( d ). Dotted lines are drawn at P  < 0.05 and black arrows indicate the stratification points used. e , An epigenetic signatured derived from CD8 + CD19.28ζ FOXO1 OE bulk ATAC-seq data was applied to pre-manufactured paediatric CAR T cell single-cell ATAC-seq data 5 . Violin plots show TCF1 OE epigenetic signature scores for patients with durable (Patient 52, n  = 616 cells; Patient 54, n  = 2959 cells) and short (Patient 38, n  = 2093 cells; Patient 66, n  = 2355 cells) CAR T cell persistence. f , GSEA was performed with CD8 + HA.28ζ TCF1 OE DEGs and DEGs derived from CD39 − CD69 − patient TILs in adult melanoma 8 . Violin plots in a , e show minima and maxima; centre lines represent mean; dashed lines represent top and bottom quartiles. Statistical comparisons were performed using two-tailed Mann–Whitney test ( a , left), log-rank Mantel–Cox test ( a , right), two-sided Wald test ( e ), and two-sided Kolmogorov–Smirnov test ( f ).

Supplementary information

Supplementary figures.

This file includes Supplementary Fig. 1 - the representative flow cytometry gating strategy, Supplementary Fig. 2 - bulk ATAC-seq quality control metrics and Supplementary Fig. 3 - raw western blot data corresponding to Extended Data Figs 6b and 8a.

Reporting Summary

Supplementary table 1.

FOXO1 regulon genes.

Supplementary Table 2

Reagents used in this study.

Peer Review File

Source data, source data fig. 1, source data fig. 2, source data fig. 3, source data fig. 4, source data fig. 5, source data extended data fig. 1, source data extended data fig. 2, source data extended data fig. 3, source data extended data fig. 4, source data extended data fig. 6, source data extended data fig. 7, source data extended data fig. 8, source data extended data fig. 9, source data extended data fig. 10, rights and permissions.

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ .

Reprints and permissions

About this article

Cite this article.

Doan, A.E., Mueller, K.P., Chen, A.Y. et al. FOXO1 is a master regulator of memory programming in CAR T cells. Nature (2024). https://doi.org/10.1038/s41586-024-07300-8

Download citation

Received : 12 April 2023

Accepted : 12 March 2024

Published : 10 April 2024

DOI : https://doi.org/10.1038/s41586-024-07300-8

Share this article

Anyone you share the following link with will be able to read this content:

Sorry, a shareable link is not currently available for this article.

Provided by the Springer Nature SharedIt content-sharing initiative

By submitting a comment you agree to abide by our Terms and Community Guidelines . If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Quick links

• Explore articles by subject
• Guide to authors
• Editorial policies

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.