presentation slides journal article

• - Google Chrome

Intended for healthcare professionals

• My email alerts
• BMA member login
• Username * Password * Forgot your log in details? Need to activate BMA Member Log In Log in via OpenAthens Log in via your institution

Search form

• Advanced search
• Search responses
• Search blogs
• How to prepare and...

How to prepare and deliver an effective oral presentation

• Related content
• Peer review
• Lucia Hartigan , registrar 1 ,
• Fionnuala Mone , fellow in maternal fetal medicine 1 ,
• Mary Higgins , consultant obstetrician 2
• 1 National Maternity Hospital, Dublin, Ireland
• 2 National Maternity Hospital, Dublin; Obstetrics and Gynaecology, Medicine and Medical Sciences, University College Dublin
• luciahartigan{at}hotmail.com

The success of an oral presentation lies in the speaker’s ability to transmit information to the audience. Lucia Hartigan and colleagues describe what they have learnt about delivering an effective scientific oral presentation from their own experiences, and their mistakes

The objective of an oral presentation is to portray large amounts of often complex information in a clear, bite sized fashion. Although some of the success lies in the content, the rest lies in the speaker’s skills in transmitting the information to the audience. 1

Preparation

It is important to be as well prepared as possible. Look at the venue in person, and find out the time allowed for your presentation and for questions, and the size of the audience and their backgrounds, which will allow the presentation to be pitched at the appropriate level.

See what the ambience and temperature are like and check that the format of your presentation is compatible with the available computer. This is particularly important when embedding videos. Before you begin, look at the video on stand-by and make sure the lights are dimmed and the speakers are functioning.

For visual aids, Microsoft PowerPoint or Apple Mac Keynote programmes are usual, although Prezi is increasing in popularity. Save the presentation on a USB stick, with email or cloud storage backup to avoid last minute disasters.

When preparing the presentation, start with an opening slide containing the title of the study, your name, and the date. Begin by addressing and thanking the audience and the organisation that has invited you to speak. Typically, the format includes background, study aims, methodology, results, strengths and weaknesses of the study, and conclusions.

If the study takes a lecturing format, consider including “any questions?” on a slide before you conclude, which will allow the audience to remember the take home messages. Ideally, the audience should remember three of the main points from the presentation. 2

Have a maximum of four short points per slide. If you can display something as a diagram, video, or a graph, use this instead of text and talk around it.

Animation is available in both Microsoft PowerPoint and the Apple Mac Keynote programme, and its use in presentations has been demonstrated to assist in the retention and recall of facts. 3 Do not overuse it, though, as it could make you appear unprofessional. If you show a video or diagram don’t just sit back—use a laser pointer to explain what is happening.

Rehearse your presentation in front of at least one person. Request feedback and amend accordingly. If possible, practise in the venue itself so things will not be unfamiliar on the day. If you appear comfortable, the audience will feel comfortable. Ask colleagues and seniors what questions they would ask and prepare responses to these questions.

It is important to dress appropriately, stand up straight, and project your voice towards the back of the room. Practise using a microphone, or any other presentation aids, in advance. If you don’t have your own presenting style, think of the style of inspirational scientific speakers you have seen and imitate it.

Try to present slides at the rate of around one slide a minute. If you talk too much, you will lose your audience’s attention. The slides or videos should be an adjunct to your presentation, so do not hide behind them, and be proud of the work you are presenting. You should avoid reading the wording on the slides, but instead talk around the content on them.

Maintain eye contact with the audience and remember to smile and pause after each comment, giving your nerves time to settle. Speak slowly and concisely, highlighting key points.

Do not assume that the audience is completely familiar with the topic you are passionate about, but don’t patronise them either. Use every presentation as an opportunity to teach, even your seniors. The information you are presenting may be new to them, but it is always important to know your audience’s background. You can then ensure you do not patronise world experts.

To maintain the audience’s attention, vary the tone and inflection of your voice. If appropriate, use humour, though you should run any comments or jokes past others beforehand and make sure they are culturally appropriate. Check every now and again that the audience is following and offer them the opportunity to ask questions.

Finishing up is the most important part, as this is when you send your take home message with the audience. Slow down, even though time is important at this stage. Conclude with the three key points from the study and leave the slide up for a further few seconds. Do not ramble on. Give the audience a chance to digest the presentation. Conclude by acknowledging those who assisted you in the study, and thank the audience and organisation. If you are presenting in North America, it is usual practice to conclude with an image of the team. If you wish to show references, insert a text box on the appropriate slide with the primary author, year, and paper, although this is not always required.

Answering questions can often feel like the most daunting part, but don’t look upon this as negative. Assume that the audience has listened and is interested in your research. Listen carefully, and if you are unsure about what someone is saying, ask for the question to be rephrased. Thank the audience member for asking the question and keep responses brief and concise. If you are unsure of the answer you can say that the questioner has raised an interesting point that you will have to investigate further. Have someone in the audience who will write down the questions for you, and remember that this is effectively free peer review.

Be proud of your achievements and try to do justice to the work that you and the rest of your group have done. You deserve to be up on that stage, so show off what you have achieved.

Competing interests: We have read and understood the BMJ Group policy on declaration of interests and declare the following interests: None.

• ↵ Rovira A, Auger C, Naidich TP. How to prepare an oral presentation and a conference. Radiologica 2013 ; 55 (suppl 1): 2 -7S. OpenUrl
• ↵ Bourne PE. Ten simple rules for making good oral presentations. PLos Comput Biol 2007 ; 3 : e77 . OpenUrl PubMed
• ↵ Naqvi SH, Mobasher F, Afzal MA, Umair M, Kohli AN, Bukhari MH. Effectiveness of teaching methods in a medical institute: perceptions of medical students to teaching aids. J Pak Med Assoc 2013 ; 63 : 859 -64. OpenUrl

• Research Process
• Manuscript Preparation
• Manuscript Review
• Publication Process
• Publication Recognition
• Language Editing Services
• Translation Services

How to Make a PowerPoint Presentation of Your Research Paper

• 4 minute read
• 138.9K views

Table of Contents

A research paper presentation is often used at conferences and in other settings where you have an opportunity to share your research, and get feedback from your colleagues. Although it may seem as simple as summarizing your research and sharing your knowledge, successful research paper PowerPoint presentation examples show us that there’s a little bit more than that involved.

In this article, we’ll highlight how to make a PowerPoint presentation from a research paper, and what to include (as well as what NOT to include). We’ll also touch on how to present a research paper at a conference.

Purpose of a Research Paper Presentation

The purpose of presenting your paper at a conference or forum is different from the purpose of conducting your research and writing up your paper. In this setting, you want to highlight your work instead of including every detail of your research. Likewise, a presentation is an excellent opportunity to get direct feedback from your colleagues in the field. But, perhaps the main reason for presenting your research is to spark interest in your work, and entice the audience to read your research paper.

So, yes, your presentation should summarize your work, but it needs to do so in a way that encourages your audience to seek out your work, and share their interest in your work with others. It’s not enough just to present your research dryly, to get information out there. More important is to encourage engagement with you, your research, and your work.

Tips for Creating Your Research Paper Presentation

In addition to basic PowerPoint presentation recommendations, which we’ll cover later in this article, think about the following when you’re putting together your research paper presentation:

• Know your audience : First and foremost, who are you presenting to? Students? Experts in your field? Potential funders? Non-experts? The truth is that your audience will probably have a bit of a mix of all of the above. So, make sure you keep that in mind as you prepare your presentation.

Know more about: Discover the Target Audience .

• Your audience is human : In other words, they may be tired, they might be wondering why they’re there, and they will, at some point, be tuning out. So, take steps to help them stay interested in your presentation. You can do that by utilizing effective visuals, summarize your conclusions early, and keep your research easy to understand.
• Running outline : It’s not IF your audience will drift off, or get lost…it’s WHEN. Keep a running outline, either within the presentation or via a handout. Use visual and verbal clues to highlight where you are in the presentation.
• Where does your research fit in? You should know of work related to your research, but you don’t have to cite every example. In addition, keep references in your presentation to the end, or in the handout. Your audience is there to hear about your work.
• Plan B : Anticipate possible questions for your presentation, and prepare slides that answer those specific questions in more detail, but have them at the END of your presentation. You can then jump to them, IF needed.

What Makes a PowerPoint Presentation Effective?

You’ve probably attended a presentation where the presenter reads off of their PowerPoint outline, word for word. Or where the presentation is busy, disorganized, or includes too much information. Here are some simple tips for creating an effective PowerPoint Presentation.

• Less is more: You want to give enough information to make your audience want to read your paper. So include details, but not too many, and avoid too many formulas and technical jargon.
• Clean and professional : Avoid excessive colors, distracting backgrounds, font changes, animations, and too many words. Instead of whole paragraphs, bullet points with just a few words to summarize and highlight are best.
• Know your real-estate : Each slide has a limited amount of space. Use it wisely. Typically one, no more than two points per slide. Balance each slide visually. Utilize illustrations when needed; not extraneously.
• Keep things visual : Remember, a PowerPoint presentation is a powerful tool to present things visually. Use visual graphs over tables and scientific illustrations over long text. Keep your visuals clean and professional, just like any text you include in your presentation.

Know more about our Scientific Illustrations Services .

Another key to an effective presentation is to practice, practice, and then practice some more. When you’re done with your PowerPoint, go through it with friends and colleagues to see if you need to add (or delete excessive) information. Double and triple check for typos and errors. Know the presentation inside and out, so when you’re in front of your audience, you’ll feel confident and comfortable.

How to Present a Research Paper

If your PowerPoint presentation is solid, and you’ve practiced your presentation, that’s half the battle. Follow the basic advice to keep your audience engaged and interested by making eye contact, encouraging questions, and presenting your information with enthusiasm.

We encourage you to read our articles on how to present a scientific journal article and tips on giving good scientific presentations .

Language Editing Plus

Improve the flow and writing of your research paper with Language Editing Plus. This service includes unlimited editing, manuscript formatting for the journal of your choice, reference check and even a customized cover letter. Learn more here , and get started today!

Know How to Structure Your PhD Thesis

Systematic Literature Review or Literature Review?

You may also like.

What is a Good H-index?

What is a Corresponding Author?

How to Submit a Paper for Publication in a Journal

Input your search keywords and press Enter.

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
• Explore content
• About the journal
• Publish with us
• Sign up for alerts
• CAREER GUIDE
• 12 May 2021

Good presentation skills benefit careers — and science

• David Rubenson 0

David Rubenson is the director of the scientific-communications firm No Bad Slides ( nobadslides.com ) in Los Angeles, California.

You can also search for this author in PubMed   Google Scholar

You have full access to this article via your institution.

A better presentation culture can save the audience and the larger scientific world valuable time and effort. Credit: Shutterstock

In my experience as a presentation coach for biomedical researchers, I have heard many complaints about talks they attend: too much detail, too many opaque visuals, too many slides, too rushed for questions and so on. Given the time scientists spend attending presentations, both in the pandemic’s virtual world and in the ‘face-to-face’ one, addressing these complaints would seem to be an important challenge.

I’m dispirited that being trained in presentation skills, or at least taking more time to prepare presentations, is often not a high priority for researchers or academic departments. Many scientists feel that time spent improving presentations detracts from research or clocking up the numbers that directly affect career advancement — such as articles published and the amount of grant funding secured. Add in the pressing, and sometimes overwhelming, bureaucratic burdens associated with working at a major biomedical research institute, and scientists can simply be too busy to think about changing the status quo.

Improving presentations can indeed be time-consuming. But there are compelling reasons for researchers to put this near the top of their to-do list.

You’re probably not as good a presenter as you think you are

Many scientists see problems in colleagues’ presentations, but not their own. Having given many lousy presentations, I know that it is all too easy to receive (and accept) plaudits; audiences want to be polite. However, this makes it difficult to get an accurate assessment of how well you have communicated your message.

Why your scientific presentation should not be adapted from a journal article

With few exceptions, biomedical research presentations are less effective than the speaker would believe. And with few exceptions, researchers have little appreciation of what makes for a good presentation. Formal training in presentation techniques (see ‘What do scientists need to learn?’) would help to alleviate these problems.

Improving a presentation can help you think about your own research

A well-designed presentation is not a ‘data dump’ or an exercise in advanced PowerPoint techniques. It is a coherent argument that can be understood by scientists in related fields. Designing a good presentation forces a researcher to step back from laboratory procedures and organize data into themes; it’s an effective way to consider your research in its entirety.

You might get insights from the audience

Overly detailed presentations typically fill a speaker’s time slot, leaving little opportunity for the audience to ask questions. A comprehensible and focused presentation should elicit probing questions and allow audience members to suggest how their tools and methods might apply to the speaker’s research question.

Many have suggested that multidisciplinary collaborations, such as with engineers and physical scientists, are essential for solving complex problems in biomedicine. Such innovative partnerships will emerge only if research is communicated clearly to a broad range of potential collaborators.

It might improve your grant writing

Many grant applications suffer from the same problem as scientific presentations — too much detail and a lack of clearly articulated themes. A well-designed presentation can be a great way to structure a compelling grant application: by working on one, you’re often able to improve the other.

It might help you speak to important, ‘less-expert’ audiences

As their career advances, it is not uncommon for scientists to increasingly have to address audiences outside their speciality. These might include department heads, deans, philanthropic foundations, individual donors, patient groups and the media. Communicating effectively with scientific colleagues is a prerequisite for reaching these audiences.

Collection: Conferences

Better presentations mean better science

An individual might not want to spend 5 hours improving their hour-long presentation, but 50 audience members might collectively waste 50 hours listening to that individual’s mediocre effort. This disparity shows that individual incentives aren’t always aligned with society’s scientific goals. An effective presentation can enhance the research and critical-thinking skills of the audience, in addition to what it does for the speaker.

What do scientists need to learn?

Formal training in scientific presentation techniques should differ significantly from programmes that stress the nuances of public speaking.

The first priority should be to master basic presentation concepts, including:

• How to build a concise scientific narrative.

• Understanding the limitations of slides and presentations.

• Understanding the audience’s time and attention-span limitations .

• Building a complementary, rather than repetitive, relationship between what the speaker says and what their slides show.

The training should then move to proper slide design, including:

• The need for each slide to have an overarching message.

• Using slide titles to help convey that message.

• Labelling graphs legibly.

• Deleting superfluous data and other information.

• Reducing those 100-word text slides to 40 words (or even less) without losing content.

• Using colour to highlight categories of information, rather than for decoration.

• Avoiding formats that have no visual message, such as data tables.

A well-crafted presentation with clearly drawn slides can turn even timid public speakers into effective science communicators.

Scientific leaders have a responsibility to provide formal training and to change incentives so that researchers spend more time improving presentations.

A dynamic presentation culture, in which every presentation is understood, fairly critiqued and useful for its audience, can only be good for science.

Nature 594 , S51-S52 (2021)

doi: https://doi.org/10.1038/d41586-021-01281-8

This is an article from the Nature Careers Community, a place for Nature readers to share their professional experiences and advice. Guest posts are encouraged .

Related Articles

• Conferences and meetings
• Research management

Science-policy advisers shape programmes that solve real-world problems

Career Feature 18 SEP 24

Can AI be used to assess research quality?

Nature Index 18 SEP 24

The grassroots organizations continuing the fight for Ukrainian science

Career Feature 11 SEP 24

Academics say flying to meetings harms the climate — but they carry on

News 13 SEP 24

Hybrid conferences should be the norm — optimize them so everyone benefits

Editorial 06 AUG 24

How to spot a predatory conference, and what science needs to do about them: a guide

Career Feature 30 JUL 24

The human costs of the research-assessment culture

Career Feature 09 SEP 24

Why I’m committed to breaking the bias in large language models

Career Guide 04 SEP 24

Binning out-of-date chemicals? Somebody think about the carbon!

Correspondence 27 AUG 24

Associate or Senior Editor, Communications Chemistry

Job Title: Associate or Senior Editor, Communications Chemistry Location(s): Berlin, New York, Jersey City or Philadelphia – Hybrid Working      Ap...

New Jersey (US)

Springer Nature Ltd

Faculty Positions in Center of Bioelectronic Medicine, School of Life Sciences, Westlake University

SLS invites applications for multiple tenure-track/tenured faculty positions at all academic ranks.

Hangzhou, Zhejiang, China

School of Life Sciences, Westlake University

Faculty Positions, Aging and Neurodegeneration, Westlake Laboratory of Life Sciences and Biomedicine

Applicants with expertise in aging and neurodegeneration and related areas are particularly encouraged to apply.

Westlake Laboratory of Life Sciences and Biomedicine (WLLSB)

Faculty Positions in Chemical Biology, Westlake University

We are seeking outstanding scientists to lead vigorous independent research programs focusing on all aspects of chemical biology including...

Faculty Positions at the Center for Machine Learning Research (CMLR), Peking University

CMLR's goal is to advance machine learning-related research across a wide range of disciplines.

Beijing, China

Center for Machine Learning Research (CMLR), Peking University

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

Quick links

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

An official website of the United States government

The .gov means it’s official. Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

The site is secure. The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

• Publications
• Account settings

The PMC website is updating on October 15, 2024. Learn More or Try it out now .

• Advanced Search
• Journal List
• Lippincott Open Access

How to deliver an oral presentation

Georgina wellstead.

a Lister Hospital, East and North Hertfordshire NHS Trust

Katharine Whitehurst

b Royal Devon and Exeter Hospital

Buket Gundogan

c University College London

d Guy's St Thomas' NHS Foundation Trust, London, UK

Delivering an oral presentation in conferences and meetings can seem daunting. However, if delivered effectively, it can be an invaluable opportunity to showcase your work in front of peers as well as receive feedback on your project. In this “How to” article, we demonstrate how one can plan and successfully deliver an engaging oral presentation.

Giving an oral presentation at a scientific conference is an almost inevitable task at some point during your medical career. The prospect of presenting your original work to colleagues and peers, however, may be intimidating, and it can be difficult to know how to approach it. Nonetheless, it is important to remember that although daunting, an oral presentation is one of the best ways to get your work out there, and so should be looked upon as an exciting and invaluable opportunity.

Slide content

Although things may vary slightly depending on the type of research you are presenting, the typical structure is as follows:

• Opening slide (title of study, authors, institutions, and date)
• Methodology
• Discussion (including strengths and weaknesses of the study)

Conclusions

Picking out only the most important findings to include in your presentation is key and will keep it concise and easy to follow. This in turn will keep your viewers engaged, and more likely to understand and remember your presentation.

Psychological analysis of PowerPoint presentations, finds that 8 psychological principles are often violated 1 . One of these was the limited capacity of working memory, which can hold 4 units of information at any 1 time in most circumstances. Hence, too many points or concepts on a slide could be detrimental to the presenter’s desire to give information.

You can also help keep your audience engaged with images, which you can talk around, rather than lots of text. Video can also be useful, for example, a surgical procedure. However, be warned that IT can let you down when you need it most and you need to have a backup plan if the video fails. It’s worth coming to the venue early and testing it and resolving issues beforehand with the AV support staff if speaking at a conference.

Slide design and layout

It is important not to clutter your slides with too much text or too many pictures. An easy way to do this is by using the 5×5 rule. This means using no more than 5 bullet points per slide, with no more than 5 words per bullet point. It is also good to break up the text-heavy slides with ones including diagrams or graphs. This can also help to convey your results in a more visual and easy-to-understand way.

It is best to keep the slide design simple, as busy backgrounds and loud color schemes are distracting. Ensure that you use a uniform font and stick to the same color scheme throughout. As a general rule, a light-colored background with dark-colored text is easier to read than light-colored text on a dark-colored background. If you can use an image instead of text, this is even better.

A systematic review study of expert opinion papers demonstrates several key recommendations on how to effectively deliver medical research presentations 2 . These include:

• Keeping your slides simple
• Knowing your audience (pitching to the right level)
• Making eye contact
• Rehearsing the presentation
• Do not read from the slides
• Limiting the number of lines per slide
• Sticking to the allotted time

You should practice your presentation before the conference, making sure that you stick to the allocated time given to you. Oral presentations are usually short (around 8–10 min maximum), and it is, therefore, easy to go under or over time if you have not rehearsed. Aiming to spend around 1 minute per slide is usually a good guide. It is useful to present to your colleagues and seniors, allowing them to ask you questions afterwards so that you can be prepared for the sort of questions you may get asked at the conference. Knowing your research inside out and reading around the subject is advisable, as there may be experts watching you at the conference with more challenging questions! Make sure you re-read your paper the day before, or on the day of the conference to refresh your memory.

It is useful to bring along handouts of your presentation for those who may be interested. Rather than printing out miniature versions of your power point slides, it is better to condense your findings into a brief word document. Not only will this be easier to read, but you will also save a lot of paper by doing this!

Delivering the presentation

Having rehearsed your presentation beforehand, the most important thing to do when you get to the conference is to keep calm and be confident. Remember that you know your own research better than anyone else in the room! Be sure to take some deep breaths and speak at an appropriate pace and volume, making good eye contact with your viewers. If there is a microphone, don’t keep turning away from it as the audience will get frustrated if your voice keeps cutting in and out. Gesturing and using pointers when appropriate can be a really useful tool, and will enable you to emphasize your important findings.

Presenting tips

• Do not hide behind the computer. Come out to the center or side and present there.
• Maintain eye contact with the audience, especially the judges.
• Remember to pause every so often.
• Don’t clutter your presentation with verbal noise such as “umm,” “like,” or “so.” You will look more slick if you avoid this.
• Rhetorical questions once in a while can be useful in maintaining the audience’s attention.

When reaching the end of your presentation, you should slow down in order to clearly convey your key points. Using phases such as “in summary” and “to conclude” often prompts those who have drifted off slightly during your presentation start paying attention again, so it is a critical time to make sure that your work is understood and remembered. Leaving up your conclusions/summary slide for a short while after stopping speaking will give the audience time to digest the information. Conclude by acknowledging any fellow authors or assistants before thanking the audience for their attention and inviting any questions (as long as you have left sufficient time).

If asked a question, firstly thank the audience member, then repeat what they have asked to the rest of the listeners in case they didn’t hear the first time. Keep your answers short and succinct, and if unsure say that the questioner has raised a good point and that you will have to look into it further. Having someone else in the audience write down the question is useful for this.

The key points to remember when preparing for an oral presentation are:

• Keep your slides simple and concise using the 5×5 rule and images.
• When appropriate; rehearse timings; prepare answers to questions; speak slowly and use gestures/ pointers where appropriate; make eye contact with the audience; emphasize your key points at the end; make acknowledgments and thank the audience; invite questions and be confident but not arrogant.

Conflicts of interest

The authors declare that they have no financial conflict of interest with regard to the content of this report.

Sponsorships or competing interests that may be relevant to content are disclosed at the end of this article.

Published online 8 June 2017

Research Communities by Springer Nature

How to make a good (and interesting) presentation in journal club.

Share this post

Choose a social network to share with, or copy the URL to share elsewhere

Share with...

...or copy the link.

communities.springernature.com

Who says science needs to be hard and dry?

When I give presentation in journal club, I always select the kind of papers that tell a "fun" story- I believe we can learn more by discussing "how the author(s) come up with such idea?" question. Over the years, the topics of my selected papers have ranged from how bugs determine the color of laid eggs to whether getting cancer is just bad luck. Many people have told me that they like how the papers I selected arouse interest and discussion from the audience in journal club. Here I'd like to explain how we all can benefit our research by reading and interpreting research papers from a different perspective.

To begin with, we have to understand that the purpose of scientific research is very different from that of scientific publication, and the latter can facilitate but does not achieve the former. Therefore, it is important for a career scientist to be able to distinguish these from each other, get to know the structural elements of both, and identify what can be learned from them for her/his own research.   

First of all, we don’t only study cancer. We study the natural history of life. Ultimately, all biological studies address different perspectives of life. Keeping in mind the quote "Nothing in biology makes sense except in the light of evolution" (Theodosius Dobzhansky, 1973), any paper is relevant.   

Second, in most studies, the authors observe the world through the lens of the contemporary paradigm or prevailing models. Many papers in top-tier journals attempt to find the “last piece” of the puzzle in the established model (and many “elite” authors are very good at this). An idea that does not fit into any of the paradigms will have a hard time getting published. A good example is Carl Woese. He single-handedly redefined the history of life but was mostly ignored until his later years because people of his time did not know where his idea should be placed (if you don’t know who he is, please Google him).

Third, we have to understand how a paper is written. Running a study is like constructing a skyscraper. You dig ground to make a foundation, lift pillars, construct floor by floor. Finally, you reach the top and finish the roof. When the construction is completed, you remove all the scaffolds and auxiliaries, clean up all the garbage, and decorate the environment. Now a brand new, beautiful building stands in front of people. But when someone asks you how such a marvelous building is constructed, you say: "I started constructing it from the roof, followed by the top floor, and floor by floor built down until the first floor touched the ground. This is a perfect plan, isn’t it?” Unfortunately, this is often how a study is presented at publication nowadays. If you follow the authors’ plans, most papers are over-decorated in a similar fashion, making them quite indistinguishable, with everything arranged perfectly and logically, even though the study hadn’t truly evolved in that way, sacrificing many critical elements that may give implications or insights to the field. For example, a discovery made by chance is described as a process following a logical design without mentioning the accident, thus the critical elements involved in the discovering circumstances may be lost forever, resulting in low reproducibility. Alternatively, following a “perfect” plan, a paper may be over-decorated with mechanism studies, and the real drivers of the phenomenon are overlooked.    

The route out of these “conceptual traps”, I believe, comes from a genuine observation or curious question that can catch people by surprise. For example, one of my all-time favorite papers is “Genetic Variations Associated with Red Hair Color and Fear of Dental Pain, Anxiety Regarding Dental Care and Avoidance of Dental Care” - yes, this is the real title. The study was initiated by an urban legend circulating among dentists: redheads have a worse response to anesthesia and terrible tooth conditions. The author - a dentist - wanted to test if it was true. What would you think if you heard such a rumor as a dentist? As you can imagine, this study is not high-profile journal material (it was published in a dental house journal, J. Am. Dental Assoc., 2009, 140:896-905). Because I study pigment cells, the results gave me a “think-out-of-the-box” moment: pigment variation and neural response are intertwined together evolutionarily. 

With these thoughts in mind, I would like to share a few “tips” for selecting a paper and preparing a presentation for journal club:

● Select a paper with a subject that might interest both scientists and non-scientists. A genuine question out of curiosity is always intriguing. Studies in lifestyle and behavior are fun because the audience can connect with them personally.

● In many cases, why the researcher asked the question and how she/he solved it are more valuable - and interesting - than the discovery itself. Even a wrong question can lead to a good observation. 

● Discuss what led the authors to the current study in the historic and/or conceptual (paradigm) perspective. This is necessary, in my opinion. For example, Joan Masague copied the in vivo cycling methodology from Isaiah J. Fidler, who got the idea from Luria-Delbruck distribution in bacterial resistance to phage. From here, we can easily see how studies of bacterial resistance heavily impacted the concept of clonal selection in cancer research. It would be very interesting to discuss the extent/limit of this concept in cancer research. Digging into the history of the research field can bring implications beyond the imagination.  

● Figure out if the question and the hypothesis are the “roof” or the “foundation” of the study. This will also arouse fun discussion.

● Examine whether the “mechanism” is required or decorative for the conclusion. Here is one of my favorite examples. In 1846, Hungarian clinician Ignaz Semmelweis published his findings in Vienna that washing deliverers’ hands with chlorinated lime solutions could effectively reduce maternal mortality in obstetrical clinics. Although the experimental data was solid, the idea was rejected by the most renowned doctors at the time, including Rudolf Virchow. The reason? Semmelweis could not offer an explanation fitting the contemporary scientific concept (i.e., “mechanism”) for his findings. The practice of hand disinfection did not prevail until Pasteur’s germ theory emerged in 1880, 15 years after Semmelweis had died in a mental institute. During this period, more women died unnecessarily because elite doctors demanded mechanisms in a scientific paper.

● Try to discuss how the findings can be applied to other fields. For example, after discovery of immune checkpoints, many immunologists tried to activate them to cure autoimmunity. Imagine this: if you read such a paper in those years, how would you think about its implication in cancer research?         

Actually, all the statements above involve only two factors: zooming out and then zooming in on the question. Believe me, doing this will easily facilitate many fun discussions.

Here are some more practical, step-by-step suggestions for the slides for journal club:

1. Start with a brief background of the field: a historic account to explain “how we got here”, and/or introduction to the current and alternative paradigms. Do these paradigms make sense in terms of biological evolution or life history? 

2. Summarize the model system and focused pathway/process being used and studied that is related to the paper. What is the scope of the model being used, and how relevant is it to the real world?

3. What is the author’s question? Why did she/he ask it? Is the question derived from the current model, or from an unexpected observation?  

4. What are the key claims in the paper? (We put this first so we can hang all the data against their claims. Ironically, the hypothesis in the paper may already give a good clue since it is often added after all the results were generated.)

5. A summary of the study design is helpful, especially for complicated projects.

6. Pick and choose key data that support the central conclusion, summarize everything else.

7. How much could the results answer the question? Alternatively, what is the paradigm-shifting discovery?

8. What is the implication of the results? How can we make use of the information in the paper in our own work? In what ways could the results impact other fields? What are the unanswered questions?

All the questions here can be asked during your presentation to arouse questions or discussion from the audience. 

Case Study: Ising C., et al. NLRP3 inflammasome activation drives tau pathology. Nature (2019).

Conclusion:  1. fibrillar amyloid-beta -> NLRP3 inflammasome -> tau kinase/phosphatase -> tau pathology  2. Neurofibrillary tangles develop downstream of amyloid-beta-induced microglial activation.

Historic context: What is the “driver” of Alzheimer disease (AD) identified by pathological and genetic studies in the history of research?

Evolutionary context: Why is there neurodegeneration disease?  1. Do other animals get neurodegeneration disease? Are the genes involved in AD conserved in other animals? What are the functions of the conserved and divergent genes? 2. “Why would we have in our brains proteins such as α-synuclein or tau that, without substantive modification appears to be able to accumulate and cause some rather distressing diseases?” (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4662249/) 3. What is the common cause of microglia activation? Is neurodegeneration disease the price we pay to prevent parasite infection in brain? 4. Does the conclusion of this study fit in any evolutionary biology explanation? If so, is the explanation supported by any epidemiological data worldwide?

Results: 1. How much in pathology can the identified mechanism explain? 2. Can boundary condition of the model be mapped to human data?

Biomedical relevance: 1. Is there any study in diet and life style related to the conclusion, so a preventive/diagnostic measure can be suggested? 2. Disease of aging and cancer are two extreme ends in the same spectrum. Is the activation of microglia relevant to the occurrence or suppression of brain tumors?

The author would like to thank Dr. Sarah Spaeh for her editorial assistance. 

Please sign in or register for FREE

If you are a registered user on Research Communities by Springer Nature, please sign in

This is an excellent article, not only about how to present but where adn what to look for. Most big findings are serendipitous, and boggles our mind. Keeping an open mind, looking for crazy connections everywhere, not just in high tier journals, is a w3onderful suggestion. Thank you

Hi, Sarawati,

I am glad that this article is helpful for you! There are so many interesting and important papers out there, not necessarily in the top-tier journals. For example, Luria-Delbruck distribution was published in the journal Gene in 1943. The work, which won them Nobel Prize, is the foundation of research in the evolution of cells, but the modern impact factor of the journal was like 4.0.

Follow the Topic

Recommended content, what is a hypothesis, is ai a hype, circle of connections, career transition in my 50s, in memory of a young postdoc.

We use cookies to ensure the functionality of our website, to personalize content and advertising, to provide social media features, and to analyze our traffic. If you allow us to do so, we also inform our social media, advertising and analysis partners about your use of our website. You can decide for yourself which categories you want to deny or allow. Please note that based on your settings not all functionalities of the site are available.

Further information can be found in our privacy policy .

Cookie Control

Customise your preferences for any tracking technology

The following allows you to customize your consent preferences for any tracking technology used to help us achieve the features and activities described below. To learn more about how these trackers help us and how they work, refer to the cookie policy. You may review and change your preferences at any time.

These trackers are used for activities that are strictly necessary to operate or deliver the service you requested from us and, therefore, do not require you to consent.

These trackers help us to deliver personalized marketing content and to operate, serve and track ads.

These trackers help us to deliver personalized marketing content to you based on your behaviour and to operate, serve and track social advertising.

These trackers help us to measure traffic and analyze your behaviour with the goal of improving our service.

These trackers help us to provide a personalized user experience by improving the quality of your preference management options, and by enabling the interaction with external networks and platforms.

• Meet the Mentors
• Get Involved
• Get the T-Shirt
• Life Science Marketing
• Community Marketing
• Custom Marketing

Join Us Sign up for our feature-packed newsletter today to ensure you get the latest expert help and advice to level up your lab work.

• Genomics & Epigenetics
• DNA / RNA Manipulation and Analysis
• Protein Expression & Analysis
• PCR & Real-time PCR
• Flow Cytometry
• Microscopy & Imaging

Cells and Model Organisms

• Analytical Chemistry and Chromatography Techniques
• Chemistry for Biologists
• Basic Lab Skills & Know-how
• Equipment Mastery & Hacks
• Managing the Scientific Literature
• Career Development and Networking
• Dealing with Fellow Scientists
• Getting Funded
• Lab Statistics & Math
• Organization & Productivity
• Personal Development
• PhD Survival
• Soft Skills & Tools
• Software & Online Tools
• Survive & Thrive
• Taming the Literature
• Writing, Publishing & Presenting
• Writing, Publishing and Presenting

Journal Club: How to Prepare Effectively and Smash Your Presentation

Journal Club. So much more than reading a paper aloud. So many ways to mess it up. Got to present one? Then read our journal club toolkit.

Published September 14, 2022

I have a Master’s Degree in Chemistry and a Ph.D. in Structural Biology. I am interested in how the shape and connectivity of molecules relate to their reactivity and function.

Journal club. It’s so much more than orally dictating a paper to your peers.

It’s an opportunity to get a bunch of intelligent people in one place to share ideas. It’s a means to expand the scientific vocabulary of you and the audience. It’s a way to stimulate inventive research design.

But there are so many ways it can go wrong.

Poorly explained papers dictated blandly to an unengaged audience. Confusing heaps of data shoehorned into long presentations. Everybody stood awkwardly outside a meeting room you thought would be free.

Equipment Mastery and Hacks

Create a Sustainable Lab with Eppendorf Biobased Consumables

From: Eppendorf

Streamline and Standardize Blood Sampling with the PAXgene Blood ccfDNA Tube (RUO)*

From: Qiagen

Whether you are unsure what journal club is, are thinking of starting one, or simply want to up your presentation game—you’ve landed on the ultimate journal club guide.

The whats, the whys, and the hows, all in one place.

What Is a Journal Club in Science?

A journal club is a series of meetings in which somebody is elected to present a research paper, its methods, and findings to a group of colleagues.

The broad goal is to stimulate discussion and ideas that the attendees may apply to their own work. Alternatively, someone may choose a paper because it’s particularly impactful or ingenious.

Usually, the presenter alternates per a rota, and attendance may be optional or compulsory.

The presenter is expected to choose, analyze, and present the paper to the attendees with accompanying slides.

The presentation is then followed by a discussion of the paper by the attendees. This is usually in the form of a series of questions and answers directed toward the presenter. Ergo , the presenter is expected to know and understand the paper and subject area to a moderate extent.

Why Have a Journal Club?

I get it. You’re a busy person. There’s a difficult research problem standing between you and your next tenure.

Why bother spending the time and energy participating in a series of meetings that don’t get you closer to achieving your scientific goals?

The answer: journal club does get you closer to achieving your scientific goals!

But it does this in indirect ways that subtly make you a better scientist. For example:

• It probably takes you out of your comfort zone .
• It makes you a better communicator.
• It makes you better at analyzing data.
• It improves your ability to critique research.
• It makes you survey relevant literature.
• It exposes you and your audience to new concepts.
• It exposes your audience to relevant literature.
• It improves the reading habits of you and your audience.
• It gets clever people talking to each other.
• It gives people a break from practical science.

It also provides a platform for people to share ideas based on their collective scientific experience. And every participant has a unique set of skills. So every participant has the potential to provide valuable insight.

This is what a good journal club should illicit.

Think of journal club as reading a book. It’s going to enrich you and add beneficially to the sum of your mental furniture, but you won’t know how until you’ve read it.

Need empirical evidence to convince you? Okay!

In 1988 a group of medical interns was split into two groups. One received journal club teaching and the other received a series of seminars. Approximately 86% of the journal club group reported improved reading habits. This compares to 0% in the group who received seminar-based teaching. [1]

Journal Club Template Structure

So now you know what journal club is, you might wonder, “how is it organized and structured?”

That’s what the rest of this article delves into. If you’re in a rush and need to head back to the lab, here’s a graphical summary (Figure 1).

Nobody likes meetings that flounder around and run over time. And while I have no data to prove it, I reckon people take less away from such meetings. Here’s a basic journal club template that assumes you are the presenter.

Introduce the Paper, Topic, Journal, and Authors

Let your audience know what you will be talking about before diving right in. Remember that repetition (of the important bits) can be a good thing.

Introducing the journal in which the paper is published will give your audience a rough idea of the prestige of the work.

And introducing the authors and their respective institutes gives your audience the option of stowing this information away and following it up with further reading in their own time.

Provide a Reason Why You Chose the Paper

Have the authors managed to circumvent sacrificing animals to achieve a goal that traditionally necessitated animal harm? Have the authors repurposed a method and applied it to a problem it’s not traditionally associated with? Is it simply a monumental feat of work and success?

People are probably more likely to listen and engage with you if they know why, in all politeness, you have chosen to use their time to talk about a given paper.

It also helps them focus on the relevant bits of your presentation and form cogent questions.

Orally Present Key Findings and Methods of the Paper

Simple. Read the paper. Understand it. Make some slides. Present.

Okay, there are a lot of ways you can get this wrong and make a hash of it. We’ll tell you how to avoid these pitfalls later on.

But for now, acknowledge that a journal club meeting starts with a presentation that sets up the main bit of it—the discussion.

Invite Your Audience to Participate in a Discussion

The discussion is the primary and arguably most beneficial component of journal club since it gives the audience a platform to share ideas. Ideas formulated by their previous experience.

And I’ve said already that these contributions are unique and have the potential to be valuable to your work.

That’s why the discussion element is important.

Their questions might concur and elaborate on the contents of the paper and your presentation of it.

Alternatively, they might disagree with the methods and/or conclusions. They might even disagree with your presentation of technical topics.

Try not to be daunted, however, as all of this ultimately adds to your knowledge, and it should all be conducted in a constructive spirit.

Summarize the Meeting and Thank Your Audience for Attending

There’s no particularly enlightening reason as to why to do these things. Summarizing helps people come away from the meeting feeling like it was a positive and rewarding thing to attend.

And thanking people for their time is a simple courtesy.

How Do You Organize It?

Basic steps if you are the organizer.

Okay, we’ve just learned what goes into speaking at the journal club. But presenter or not, the responsibility of organizing it might fall to you.

So, logistically , how do you prepare a journal club? Simply follow these 5 steps:

• Distribute copies of the research article to potential participants.
• Arrange a meeting time and location.
• Organize a speaker.
• Hold the journal club.
• Seek feedback on the quality of the meeting.

Apart from point 5, these are fairly self-explanatory. Regarding point 5, feedback is essential to growing as a scientist and presenter. The easiest way to seek feedback is simply to ask.

Alternatively, you could create a form for all the meetings in the series and ask the audience to complete and return it to you.

Basic Steps If You Are the Speaker

If somebody has done all the logistics for you, great! Don’t get complacent, however.

Why not use the time to elevate your presentation to make your journal club contribution memorable and beneficial?

Don’t worry about the “hows” because we’re going to elaborate on these points, but here are 5 things you can do to ace your presentation:

• Don’t leave it to the last minute.
• Know your audience.
• Keep your presentation slides simple.
• Keep your audience engaged.
• Be open to questions and critiques.

Regarding point 1, giving yourself sufficient time to thoroughly read the article you have chosen to present ensures you are familiar with the material in it. This is essential because you will be asked questions about it. A confident reply is the foundation of an enlightening discussion.

Regarding point 3, we’re going to tell you exactly how to prepare effective slides in its own section later. But if you are in a rush, minimize the use of excessive text. And if you provide background information, stick to diagrams that give an overview of results from previous work. Remember: a picture speaks louder than a thousand words.

Regarding point 4, engagement is critical. So carry out a practice run to make sure you are happy with the flow of your presentation and to give you an idea of your timing. It is important to stick to the time that is allotted for you.

This provides good practice for more formal conference settings where you will be stopped if you run over time. It’s also good manners and shows consideration for the attendees.

And regarding point 5, as the presenter, questions are likely to be directed toward you. So anticipate questions from the outset and prepare for the obvious ones to the best of your ability.

There’s a limit to everyone’s knowledge, but being unable to provide any sort of response will be embarrassing and make you seem unprepared.

Anticipate that people might also disagree with any definitions you make and even with your presentation of other people’s data. Whether or not you agree is a different matter, but present your reasons in a calm and professional manner.

If someone is rude, don’t rise to it and respond calmly and courteously. This shouldn’t happen too often, but we all have “those people” around us.

How Do You Choose a Journal Club Paper?

Consider the quality of the journal.

Just to be clear, I don’t mean the paper itself but the journal it’s published in.

An obscure journal is more likely to contain science that’s either boring, sloppy, wrong, or all three.

And people are giving up their time and hope to be stimulated. So oblige them!

Journal impact factor and rejection rate (the ratio of accepted to rejected articles) can help you decide whether a paper is worth discussing.

Consider the Impact and Scope of the Paper

Similar to the above, but remember, dross gets published in high-impact journals too. Hopefully, you’ve read the paper you want to present. But ask yourself what makes this particular paper stand out from the millions of others to be worth presenting.

Keep It Relevant and Keep It Interesting

When choosing a paper to present, keep your audience in mind. Choose something that is relevant to the particular group you are presenting to. If only you and a few other people understand the topic, it can come off as elitist.

How Do You Break Down and Present the Paper?

Know and provide the background material.

Before you dive into the data, spend a few minutes talking about the context of the paper. What did the authors know before they started this work? How did they formulate their hypothesis? Why did they choose to address it in this way?

You may want to reference an earlier paper from the same group if the paper represents a continuation of it, but keep it brief.

Try to explain how this paper tackles an unanswered question in the field.

Understand the Hypothesis and Methods of the Paper

Make a point of stating the  hypothesis  or  main question  of the paper, so everyone understands the goal of the study and has a foundation for the presentation and discussion.

Everyone needs to start on the same foot and remain on the same page as the meeting progresses.

Turn the Paper into a Progression of Scientific Questions

Present the data as a logical series of questions and answers. A well-written paper will already have done the hard work for you. It will be organized carefully so that each figure answers a specific question, and each new question builds on the answer from the previous figure.

If you’re having trouble grasping the flow of the paper, try writing up a brief outline of the main points. Try putting the experiments and conclusions in your own words, too.

Feel free to leave out parts of the figures that you think are unnecessary, or pull extra data from the supplemental figures if it will help you explain the paper better.

Ask Yourself Questions about the Paper Before You Present

We’ve touched on this already. This is to prepare you for any questions that are likely to be asked of you. When you read the paper, what bits didn’t you understand?

Simplify Unfamiliar and Difficult Concepts

Not everyone will be familiar with the same concepts. For example, most biologists will not have a rigorous definition of entropy committed to memory or know its units. The concept of entropy might crop up in a biophysics paper, however.

Put yourself in the audience’s shoes and anticipate what they might not fully understand given their respective backgrounds.

If you are unsure, ask them if they need a definition or include a short definition in your slides.

Sum Up Important Conclusions

After you’ve finished explaining the nitty-gritty details of the paper, conclude your presentation of the data with a list of significant findings.

Every conclusion will tie in directly to proving the major conclusion of the paper. It should be clear at this point how the data answers the main question.

How Do You Present a Journal Club Powerpoint?

Okay, so we’ve just gone through the steps required to break down a paper to present it effectively at journal club. But this needs to be paired with a PowerPoint presentation, and the two bridged orally by your talk. How do you ace this?

Provide Broad Context to the Research

We are all bogged down by minutia and reagents out of necessity.

Being bogged down is research. But it helps to come up for air. Ultimately, how will the research you are about to discuss benefit the Earth and its inhabitants when said research is translated into actual products?

Science can be for its own sake, but funded science rarely is. Reminding the journal club audience of the widest aims of the nominated field provides a clear starting point for the discussion and shows that you understand the efficacy of the research at its most basic level.

The Golden Rule: A Slide per Minute

Remember during lectures when the lecturer would open PowerPoint, and you would see, with dismay, that their slides went up to 90 or something daft? Then the last 20 get rushed through, but that’s what the exam question ends up being based on.

Don’t be that person!

A 10-15 minute talk should be accompanied by? 10-15 slides! Less is more.

Be Judicious about the Information You Choose to Present

If you are present everything in the paper, people might as well just read it in their own time, and we can call journal club off.

Try to abstract only the key findings. Sometimes technical data is necessary for what you are speaking about because their value affects the efficacy of the data and validity of the conclusions.

Most of the time, however, the exact experimental conditions can be left out and given on request. It’s good practice to put all the technical data that you anticipate being asked for in a few slides at the end of your talk.

Use your judgment.

Keep the Amount of Information per Slide Low for Clarity

Your audience is already listening to you and looking at the slides, so they have a limited capacity for what they can absorb. Overwhelming them with visual queues and talking to them will disengage them.

Have only a few clearly related images that apply directly to what you speaking about at the time. Annotate them with the only key facts from your talk and develop the bigger picture verbally.

This will be hard at first because you must be on the ball and confident with your subject area and speaking to an audience.

And definitely use circles, boxes, and arrows to highlight important parts of figures, and add a flowchart or diagram to explain an unfamiliar method.

Keep It Short Overall

The exact length of your meeting is up to you or the organizer. A 15-minute talk followed by a 30-minute discussion is about the right length, Add in tea and coffee and hellos, and you get to an hour.

We tend to speak at 125-150 words per minute. All these words should not be on your slides, however. So, commit a rough script to memory and rehearse it.

You’ll find that the main points you need to mention start to stand out and fall into place naturally. Plus, your slides will serve as visual queue cards.

How Do You Ask a Question in Journal Club?

A well-organized journal club will have clear expectations of whether or not questions should be asked only during the discussion, or whether interruptions during the presentation are allowed.

And I don’t mean literally how do you soliloquize, but rather how do you get an effective discussion going.

Presenters: Ask Questions to the Audience

We all know how it goes. “Any questions?” Silence.

Scientists, by their very nature, are usually introverted. Any ideas they might want to contribute to a discussion are typically outweighed by the fear of looking silly in front of their peers. Or they think everyone already knows the item they wish to contribute. Or don’t want to be publicly disproven. And so on.

Prepare some questions to ask the audience in advance. As soon as a few people speak, everyone tends to loosen up. Take advantage of this.

Audience: Think About Topics to Praise or Critique

Aside from seeking clarification on any unclear topics, you could ask questions on:

• Does the data support the conclusions?
• Are the conclusions relevant?
• Are the methods valid?
• What are the drawbacks and limitations of the conclusions?
• Are there better methods to test the hypothesis?
• How will the research be translated into real-world benefits?
• Are there obvious follow-up experiments?
• How well is the burden of proof met?
• Is the data physiologically relevant?
• Do you agree with the conclusions?

How to Keep It Fun

Make it interactive.

Quizzes and polls are a great way to do this! And QR codes make it really easy to do on-the-fly. Remember, scientists, are shy. So why not seek their participation in an anonymized form?

You could poll your audience on the quality of the work. You could make a fun quiz based on the material you’ve covered. You could do a live “what happened next?” You could even get your feedback this way. Here’s what to do:

• Create your quiz or poll using Google forms .
• Make a shareable link.
• Paste the link into a free QR code generator .
• Put the QR code in the appropriate bit of your talk.

Use Multimedia

Talking to your audience without anything to break it up is a guaranteed way of sending them all to sleep.

Consider embedding demonstration videos and animations in your talk. Or even just pausing to interject with your own anecdotes will keep everyone concentrated on you.

Keep It Informal

At the end of the day, we’re all scientists. Perhaps at different stages of our careers, but we’ve all had similar-ish trajectories. So there’s no need for haughtiness.

And research institutes are usually aggressively casual in terms of dress code, coffee breaks, and impromptu chats. Asking everyone to don a suit won’t add any value to a journal club.

Your Journal Club Toolkit in Summary

Anyone can read a paper, but the value lies in understanding it and applying it to your own research and thought process.

Remember, journal club is about extracting wisdom from your colleagues in the form of a discussion while disseminating wisdom to them in a digestible format.

Need some inspiration for your journal club? Check out the online repositories hosted by PNAS and NASPAG to get your juices flowing.

We’ve covered a lot of information, from parsing papers to organizational logistics, and effective presentation. So why not bookmark this page so you can come back to it all when it’s your turn to present?

While you’re here, why not ensure you’re always prepared for your next journal club and download bitesize bio’s free journal club checklist ?

And if you present at journal club and realize we’ve left something obvious out. Get in touch and let us know. We’ll add it to the article!

• Linzer M et al . (1988) Impact of a medical journal club on house-staff reading habits, knowledge, and critical appraisal skills . JAMA 260 :2537–41

Share this article:

More 'Writing, Publishing and Presenting' articles

An Introduction to Biosafety Levels

Biological agents, including bacteria, viruses, parasites, and fungi, are categorized into four biosafety levels (BSL), each indicating a different threat level. These levels increase progressively, with level one organisms posing the least risk and level four the greatest. This article details each biosafety level, explaining their necessity and providing examples of organisms handled at each level.

Tissue Processing For Histology: What Exactly Happens?

Tissue processing for histology is a key step between fixation and embedding. We take you through the steps of tissue processing in this simple guide.

How to Handle Bases in the Lab: Everything You Need to Know

We all know acids are dangerous chemicals, and you should handle them carefully. But how should we handle bases? Read on for top tips for handling bases safely in the lab.

What Is Reproducible Research?

Once upon a time, I thought reproducible research meant if someone else showed X in a paper, then I should be able to get X in my experiment. However, this actually refers to replication, an important but separate concept. Reproducible research is data analysis that starts with the raw data and arrives at the same…

Scientific Illustrations Part II: Molecular Graphics and Animation

How do you explain to your friends and family what you have you’ve been working on all this time? Conveying the true wonder and intricacy of your work to the layperson can be tricky. They won’t be familiar with the lingo, and they might not immediately know how to interpret scientific graphs and figures. You…

How to Create an Effective PowerPoint Presentation

Presenting your work is a fantastic opportunity to get feedback on your project, demonstrate the significance of your results, and make the connections that will enhance your future career. And yet, how many incomprehensible lab meetings have we all sat through? How many seminars have you attended that left you feeling more confused than inspired?…

We collate wisdom and tools from researchers worldwide to help you to accelerate your progress.

Sign up now to get it in your inbox

Newsletters

• Technical Skills
• More Skills
• Microscopy Focus
• Privacy Policy
• Cookie Policy
• Terms of Use

Copyright © Science Squared – all rights reserved

10 Things Every Molecular Biologist Should Know

The eBook with top tips from our Researcher community.

Prepare For Journal Club

• Find Articles for Journal Club
• How to Evaluate Articles

Presentation Skills

Journal club examples.

• Start Searching Here
• 18 Tips for Killer Presentations - Lifehack General tips for presenting successfully.
• Creating Your 20.109 [Journal Club] Presentation - Atissa Banuazizi, MIT PowerPoint with basic tips for planning and presenting.
• How to Give a Dynamic Scientific Presentation - Elsevier Connect General tips on preparing the presentation, presenting, and PowerPoint slides.
• How to Give a Great Scientific Talk - Nature General tips on how to present or give a talk.
• Presentation Skills - Sheffield Hallam University Library Guide with tips for planning and organizing presentations and coping with nerves.
• Presenting with Poise - University of Saskatchewan PDF guide on presentation skills from the Student Learning Services at the University of Saskatchewan
• Example Journal Club Presentation - MERIT-HF This example PowerPoint presentation slide set is for the article: MERIT-HF Study Group. Effect of metoprolol CR/XL in chronic heart failure (MERIT-HF). Lancet. 1999;353:2001-2007.
• Journal Club Example Handout - MERIT-HF This example handout is for the article: MERIT-HF Study Group. Effect of metoprolol CR/XL in chronic heart failure (MERIT-HF). Lancet. 1999;353:2001-2007.
• Nursing Times Journal Club Example Handout This example handout is for the article: Mannix K, Jones C (2020) Nurses’ experiences of transitioning into advanced practice roles. Nursing Times [online]; 116: 3, 35-38.

Proper citations are an essential component of a good journal club presentation.

• Citing AMA Style 11th ed. Quick guide of rules and examples for citing and references in the AMA 11th Edition.
• APA Style 7th ed. Quick Guide Common Reference Examples Guide for APA 7th edition
• Cite & Write The library's guide to formatting and citation in AMA or APA style as well as writing skills and grammar.
• EndNote Guide EndNote is a powerful tool for managing and using your citations (references), articles, and research. With this tool, you can collect and create citations to insert directly into your Word documents.
• << Previous: How to Evaluate Articles
• Next: Start Searching Here >>
• Last Updated: Sep 19, 2024 11:08 AM
• URL: https://infoguides.roseman.edu/journalclub

AI Presentation Generator

AI Presentation Maker

AI Image Generator

Word to PPT

Reports to PPT

Outline to PPT

Research Papers to PPT

YouTube Video to PPT

PPT to Word

PPT to Text

AI PDF Summarizer

AI Word Summarizer

AI File Summarizer

AI Document Summarizer

Convert to PPT

Convert from PPT

AI Summarizer

Convert Journals to PPT with AI

Summarize a Journal into a PowerPoint Presentation for quick review

Select and upload a Journal that needs to be summarized for a presentation.

Choose from a variety of presentation template styles and select the one that best represents your content.

Relax and Watch the Magic Happen. Sit back and let AI do the heavy lifting for you! Get a customized design and stunning presentation filled with informative and professional content.

You can then edit the presentation using your preferred application, such as MS PowerPoint or Google Slides, or with our online AI Presentation Maker.

Superfast presentation creation

Join 1 million professionals, students, and educators

✓ Create with AI        ✓ Convert to PPT with AI        ✓ Compatible with PowerPoint        ✓ Built in templates        ✓ Auto Layout

Got any suggestions?

We want to hear from you! Send us a message and help improve Slidesgo

Top searches

Trending searches

mexican independence

54 templates

hispanic heritage month

21 templates

49 templates

22 templates

independencia de mexico

14 templates

indigenous canada

47 templates

Journal Presentation templates

Extra, extra the slidesgo journal is back with more news that will fill your day with knowledge these google slides & powerpoint templates about journals will bring your content to the headline after you edit them. what is happening now at the world share the best themes with your readers with cool, creative presentations.

• Calendar & Weather
• Infographics
• Marketing Plan
• Project Proposal
• Social Media
• Thesis Defense
• Black & White
• Craft & Notebook
• Floral & Plants
• Illustration
• Interactive & Animated
• Professional
• Instagram Post
• Instagram Stories

It seems that you like this template!

Register for free and start downloading now

History of journalism with whiteboard style.

Who said journalism is no longer necessary? Prove the opposite by reviewing its entire history. Highlight the most important moments, how it started and what changes it has undergone until today. You can help yourself with this creative black and white template. Its doodle style will immediately capture the attention...

Journalism Workshop: How to Create a Report

Download the Journalism Workshop: How to Create a Report presentation for PowerPoint or Google Slides. If you are planning your next workshop and looking for ways to make it memorable for your audience, don’t go anywhere. Because this creative template is just what you need! With its visually stunning design,...

Journalist Portfolio

Download the "Journalist Portfolio" presentation for PowerPoint or Google Slides. When a potential client or employer flips through the pages of your portfolio, they're not just looking at your work; they're trying to get a sense of who you are as a person. That's why it's crucial to curate your...

Editorial Meeting

Is your team composed by amazing writers, tenacious journalists and creative photographers? Our guess is that you’re an editorial! Well, if you want to have a meeting with your team members, why not make it in a creative way that reflects their personality? With this design for Google Slides and...

Premium template

Unlock this template and gain unlimited access

Journalism & Broadcasting Marketing Plan

Download the Journalism & Broadcasting Marketing Plan presentation for PowerPoint or Google Slides. This incredible template is designed to help you create your own marketing plan that is sure to impress your entire team. Using this amazing tool, you'll be able to analyze your target audience, assess your competitors, map...

Newspaper Clipping Style for History Class Infographics

Extra, extra! The latest template that compiles around 30 infographic designs hits the market! Newspapers have been spreading news for decades and decades. Those printed in paper might be in the decline nowadays compared to their digital counterparts, but we still believe! This template combines digital (it's for Google Slides...

This is My Life Project

Download the This is My Life Project presentation for PowerPoint or Google Slides and start impressing your audience with a creative and original design. Slidesgo templates like this one here offer the possibility to convey a concept, idea or topic in a clear, concise and visual way, by using different...

Funny Digital Journal for Class

Are you one of those who tend to forget assignments easily? That won't be a problem anymore thanks to this digital classroom journal! Its grid background emulates a notebook, and we've included resources for you to write down your schedules, homework, notes, important dates, and more. Plus, it's interactive! You...

Newspaper Headline Style Newsletter

You have exciting news to share with all your subscribers! These news are so big, they would fit in an entire newspaper... Well, how about doing exactly that? Our latest template for newsletters mimics a bit the style that this kind of periodical publication has, but a little more stylized...

My Internship Journal

Some degrees require you to make an internship journal after finishing your stay at a company, any many times it counts as a part of the final grade. Why not make it as a presentation? Creativity and design always make a good impression, and the different resources we have included...

Formal Digital Journal

"Dear diary, today I've been modifying twelve pages of a calendar that I found on the street. It had the month of February with 31 days! But I'm fine!" This won't happen again because with this template, which is totally editable, you'll get a journal or a diary with a...

Newspaper Clipping Style for History Class

What better way to spread knowledge about history than with a newspaper design? With this vintage style template, you'll be able to teach an amazing history lesson. It's multipurpose, so you can adapt it to the school level you want and about the historical moment you want. Did you know...

Daily Journal Infographics

Being methodical at work is essential to be successful and meet all the goals you set for yourself. It is always said that there's more than one way to skin a cat, but if you want to share yours with the rest, this daily journal infographics template is perfect for...

Writing a News Article Workshop

Hey there, news writer extraordinaire! Are you ready to spice up your workshop? This free Google Slides and PowerPoint template is the perfect tool to help you facilitate an engaging and informative news article writing workshop. Help your participants master the art of news article writing with a user-friendly interface,...

Communications Major for College: Print Journalism

Newspapers never go out of style, so if you like journalism you might be interested in this area of communication: print journalism! With this formal template of gray and red slides you can speak about the content of this interesting major like what it is about, how it is organized,...

Communications Major for College: Broadcast Journalism

Do you want to become the best communicator ever seen? With this creative design full of modern illustrations and editable resources you can speak about broadcast journalism, the new wave of journalism that has now overcome traditional media. How does this work? What’s the best platform to share news and...

Printable Top Secret Journal

The secrets that lie inside these slides are only to be read by those who have the key! Keeping a diary is a great way of organizing your thoughts and having a place to write down what goes on in your life, what went on at school that day, how...

Slideshow for Journal Clubs

Download the Slideshow for Journal Clubs presentation for PowerPoint or Google Slides and start impressing your audience with a creative and original design. Slidesgo templates like this one here offer the possibility to convey a concept, idea or topic in a clear, concise and visual way, by using different graphic...

• Page 1 of 4

Click through the PLOS taxonomy to find articles in your field.

For more information about PLOS Subject Areas, click here .

Loading metrics

Open Access

Peer-reviewed

Research Article

CBL mutations in chronic myelomonocytic leukemia often occur in the RING domain with multiple subclones per patient: Implications for targeting

Roles Conceptualization, Formal analysis, Methodology, Resources, Writing – original draft, Writing – review & editing

Affiliations Discipline of Medicine, Adelaide Medical School, Faculty of Health and Medical Sciences, The University of Adelaide, Adelaide, SA, Australia, Precision Cancer Medicine Theme, South Australian Health and Medical Research Institute (SAHMRI), University of Adelaide, Adelaide, SA, Australia

Roles Formal analysis, Writing – review & editing

Affiliation Cytokine Receptor Laboratory, SA Pathology, Adelaide, SA, Australia

Roles Formal analysis, Investigation, Methodology, Writing – review & editing

Affiliations Discipline of Medicine, Adelaide Medical School, Faculty of Health and Medical Sciences, The University of Adelaide, Adelaide, SA, Australia, Precision Cancer Medicine Theme, South Australian Health and Medical Research Institute (SAHMRI), University of Adelaide, Adelaide, SA, Australia, College of Medicine and Public Health, Flinders Health and Medical Research Institute, Flinders University, Adelaide, SA, Australia

Roles Methodology

Roles Funding acquisition, Resources, Writing – review & editing

Affiliations Discipline of Medicine, Adelaide Medical School, Faculty of Health and Medical Sciences, The University of Adelaide, Adelaide, SA, Australia, Cytokine Receptor Laboratory, SA Pathology, Adelaide, SA, Australia

Roles Funding acquisition

Roles Funding acquisition, Writing – review & editing

Affiliations Discipline of Medicine, Adelaide Medical School, Faculty of Health and Medical Sciences, The University of Adelaide, Adelaide, SA, Australia, Precision Cancer Medicine Theme, South Australian Health and Medical Research Institute (SAHMRI), University of Adelaide, Adelaide, SA, Australia, College of Medicine and Public Health, Flinders Health and Medical Research Institute, Flinders University, Adelaide, SA, Australia, SA Pathology, Adelaide, SA, Australia, Department of Hematology and Bone Marrow Transplantation, Royal Adelaide Hospital, Adelaide, SA, Australia

Affiliation QIMR Berghofer Medical Research Institute, Brisbane, QLD, Australia

Affiliation Austin Health, Heidelberg, VIC, Australia

Affiliation SA Pathology, Adelaide, SA, Australia

Affiliations Discipline of Medicine, Adelaide Medical School, Faculty of Health and Medical Sciences, The University of Adelaide, Adelaide, SA, Australia, Precision Cancer Medicine Theme, South Australian Health and Medical Research Institute (SAHMRI), University of Adelaide, Adelaide, SA, Australia, Royal Perth Hospital, Perth, WA, Australia, The University of Western Australia Medical School, Perth, WA, Australia

Affiliations Precision Cancer Medicine Theme, South Australian Health and Medical Research Institute (SAHMRI), University of Adelaide, Adelaide, SA, Australia, Department of Hematology and Bone Marrow Transplantation, Royal Adelaide Hospital, Adelaide, SA, Australia

Affiliation Precision Cancer Medicine Theme, South Australian Health and Medical Research Institute (SAHMRI), University of Adelaide, Adelaide, SA, Australia

Affiliations Precision Cancer Medicine Theme, South Australian Health and Medical Research Institute (SAHMRI), University of Adelaide, Adelaide, SA, Australia, SA Pathology, Adelaide, SA, Australia, Department of Hematology and Bone Marrow Transplantation, Royal Adelaide Hospital, Adelaide, SA, Australia

Roles Conceptualization, Funding acquisition, Methodology, Supervision, Writing – review & editing

* E-mail: [email protected]

Affiliations Discipline of Medicine, Adelaide Medical School, Faculty of Health and Medical Sciences, The University of Adelaide, Adelaide, SA, Australia, Precision Cancer Medicine Theme, South Australian Health and Medical Research Institute (SAHMRI), University of Adelaide, Adelaide, SA, Australia, SA Pathology, Adelaide, SA, Australia, Department of Hematology and Bone Marrow Transplantation, Royal Adelaide Hospital, Adelaide, SA, Australia

• Kelly Lim, 
• Winnie L. Kan, 
• Pramod C. Nair, 
• Monika Kutyna, 
• Angel F. Lopez, 
• Timothy Hercus, 
• David M. Ross, 
• Steven Lane, 
• Chun Yew Fong, 

• Published: September 19, 2024
• https://doi.org/10.1371/journal.pone.0310641
• Reader Comments

Chronic myelomonocytic leukemia (CMML) is a rare blood cancer of older adults (3 in every 1,000,000 persons) characterized by poor survival and lacking effective mutation-specific therapy. Mutations in the ubiquitin ligase Cbl occur frequently in CMML and share biological and molecular features with a clonal disease occurring in children, juvenile myelomonocytic leukemia (JMML). Here we analyzed the clinical presentations, molecular features and immunophenotype of CMML patients with CBL mutations enrolled in a prospective Phase II clinical trial stratified according to molecular markers. Clinically, CBL mutations were associated with increased bone marrow blasts at diagnosis, leukocytosis and splenomegaly, similar to patients harboring NRAS or KRAS mutations. Interestingly, 64% of patients presented with more than one CBL variant implying a complex subclonal architecture, often with co-occurrence of TET2 mutations. We found CBL mutations in CMML frequently clustered in the RING domain in contrast to JMML, where mutations frequently involve the linker helix region ( P <0.0001). According to our comparative alignment of available X-ray structures, mutations in the linker helix region such as Y371E give rise to conformational differences that could be exploited by targeted therapy approaches. Furthermore, we noted an increased percentage of CMML CD34 + stem and progenitor cells expressing CD116 and CD131 in all CBL mutant cases and increased CD116 receptor density compared to healthy controls, similar to CMML overall. In summary, our data demonstrate that CBL mutations are associated with distinct molecular and clinical features in CMML and are potentially targetable with CD116-directed immunotherapy.

Citation: Lim K, Kan WL, Nair PC, Kutyna M, Lopez AF, Hercus T, et al. (2024) CBL mutations in chronic myelomonocytic leukemia often occur in the RING domain with multiple subclones per patient: Implications for targeting. PLoS ONE 19(9): e0310641. https://doi.org/10.1371/journal.pone.0310641

Editor: Michael Massiah, George Washington University, UNITED STATES OF AMERICA

Received: April 11, 2024; Accepted: August 30, 2024; Published: September 19, 2024

Copyright: © 2024 Lim et al. This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Data Availability: All relevant data are within the manuscript and its Supporting Information files.

Funding: D. Thomas is supported by a Commonwealth Serum Laboratories (2020 Centenary Fellowship), Australian Medical Research Future Fund (1201012, 2024427, 2008972) and National Health and Medical Research Council (2023/GNT2029809) and The Leukemia and Lymphoma Society (LLS 6619-21; LLS 6650-23; CMML Special Initiative) ( www.lls.org ), Snowdome Foundation (co-funder LLS 6619-21, LLS 6650-23) ( www.snowdome.org.au ) and the Leukaemia Foundation (co-funder LLS 6619-21, LLS 6650-23) ( www.leukaemia.org.au ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing interests: The authors have declared that no competing interests exist.

Introduction

Monocytes give rise to tissue macrophages that can perform a myriad of biological functions ranging from innate immune activation to phagocytosis and wound healing. Chronic myelomonocytic leukemia (CMML) is a rare blood cancer of older adults (3 in every 1,000,000 persons) [ 1 , 2 ] characterized by an increase in clonal CD14 + CD16 – classical monocytes and their precursors in the blood and bone marrow [ 3 , 4 ]. Because of its subtle presentation, the disease is often diagnosed late, but is likely to rise in prevalence due to routine uptake of next-generation sequencing, lower threshold monocytosis criteria by the World Health Organization and increasing recognition by physicians, especially in persons previously treated with cytotoxic therapy (therapy-related CMML) [ 5 , 6 ]. Many patients present with autoinflammatory features such as vasculitis, polychondritis, Sweet syndrome and pleural/pericardial effusions [ 7 – 10 ] but the mechanism and intersection of clonal monocytes and innate vs . adaptive immunity is not understood.

The molecular pathogenesis of CMML is only beginning to be characterized. Though 70% of CMML patients present without any cytogenetic abnormalities [ 11 , 12 ] they harbor somatic mutations in genes that influence epigenetic regulation ( TET2 , DNMT3A , ASXL1 , EZH2 , IDH1 , IDH2 ), mRNA splicing ( SRSF2 , U2AF1 ), genome stability ( SETBP1 , TP53 ), transcription regulation ( RUNX1 , CEBPA , NPM1 ) and cell signaling pathways ( KRAS , NRAS , CBL , PTPN11 , JAK2 , MPL ) [ 13 – 17 ]. Interestingly, CMML shares some biological and morphological features with a clonal disease occurring in young children, juvenile myelomonocytic leukemia (JMML). Around 90% of cases of JMML are associated with mutations in the RAS signaling pathway ( PTPN11 , NRAS , KRAS , NF1 and CBL ) [ 18 – 21 ]. Notably, while the overall molecular patterns of CMML and JMML are distinct, mutations in CBL are found with equal frequency, at approximately 15% [ 17 , 18 , 22 , 23 ], in both diseases. Both CMML and JMML display hypersensitivity to the pro-inflammatory cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) which promote the differentiation of classical pro-inflammatory monocytes [ 24 , 25 ]. Emerging reports suggest that CBL mutations are associated with inferior survival in both CMML and JMML [ 15 , 26 ].

The CBL gene is located on 11q23.3 and encodes an E3 ubiquitin ligase (c-Cbl or Cbl) that acts as both a positive and negative regulator in the signal transduction of activated receptor tyrosine kinases (RTKs) and cytokine receptors. Cbl relays signals downstream of activated RTKs by functioning as an adaptor [ 27 – 30 ], and at the same time, attenuates signaling by promoting the ubiquitination of RTKs through its E3 ligase activity, marking them for degradation by the proteasome or via endocytosis [ 31 – 33 ]. Cbl recognizes phosphorylated tyrosines on active RTKs through its Src homology 2 domain within the tyrosine kinase binding domain (TKBD), and binds E2 ubiquitin-conjugating enzymes though its conserved RING domain. The TKBD and RING domain are connected by a 28 amino acid sequence, referred to as the linker helix region (LHR). A proline-rich region, serine-rich region, several C-terminal principal phosphorylation sites (Y674, Y700, Y731, Y774) and a ubiquitin-association domain (UBA) complete the structure of Cbl. Several studies on the role of the LHR and RING domain have shown that mutation of these domains, could lead to loss of activity and/or gain of oncogenicity [ 34 – 42 ].

To date, there has not been significant mutation-specific therapy developed for CMML, unlike chronic myeloid leukemia, and standard-of-care with hypomethylating agents azacitidine or decitabine is not curative [ 16 , 43 – 47 ]. While allogeneic hematopoietic stem cell transplantation may be potentially curative, stem cell transplantation is not a viable option for most older CMML patients [ 48 , 49 ]. Currently, survival is estimated at a median of 31 months, with even shorter life expectancy for patients with CMML-2 (more than 10% bone marrow blasts/promonocytes). Transformation to acute myeloid leukemia (AML), an aggressive cancer with poor long-term survival, occurs in up to 20% of CMML patients within 5 years [ 50 – 52 ].

In this study, we analyzed the clinical presentation of CMML patients with CBL mutations enrolled in a prospective clinical trial. Strikingly, 7 out of 11 (64%) patients were found to have more than one CBL clone, implying a complex clonal architecture. Clinically, CBL mutations were associated with a more proliferative phenotype evidenced by increased bone marrow blasts, leukocytosis and splenomegaly, similar to other RAS pathway mutations such as KRAS , NRAS and PTPN11 . We also found that CMML CBL mutations often co-occurred with TET2 mutations and were enriched in the RING domain compared to the LHR ( P <0.0001). Furthermore, we noted an increased percentage of CD116 and CD131-expressing CMML CD34 + progenitors compared to healthy controls. In summary, our data suggest that CBL mutants are associated with distinct clinical and molecular features in CMML.

Materials and methods

Between 1 October 2021 and 30 September 2023, 24 patients with CMML, diagnosed according to the 2016 WHO Classification of Myeloid Neoplasms [ 53 ], who met the eligibility criteria (untreated CMML with high white cell count, cytopenia or constitutional symptoms) were enrolled in the trial with written, informed consent. Detection of TET2 , KRAS , NRAS or CBL mutation at a variant allele frequency (VAF) percentage of ≥3% was a key inclusion criteria. Of these patients, 13 were male and 11 were female. The median age was 73 years (range 56−86 years). Written informed consent for genetic analysis and use of laboratory results and samples for scientific research were obtained during trial enrolment. The trial was approved in multiple centers across Australia including the Royal Adelaide Hospital, Royal Brisbane and Women’s Hospital and Austin Health. The trial was conducted with approval from the Central Adelaide Local Health Network Human Research Ethics Committee (2021/HRE00017) and registered on the Australian and New Zealand Clinical Trials Registry (ANZCTR) (Registration number ACTRN12621000223831, Acronym PREACH-M).

Clinical presentation

All clinical and laboratory data were acquired at screening, prior to commencement of the therapy protocol outlined in the trial. Complete blood examination and bone marrow (BM) analyses were performed for each patient. The spleen craniocaudal length was determined by ultrasonography.

Mutation screening

Targeted enrichment of selected coding exons and flanking intronic regions of 46 genes was performed using a custom-designed hematological neoplasms capture panel (Integrated DNA Technologies; HaemV1) and analyzed by next-generation sequencing (NGS) (Illumina NextSeq sequencing system). Variant calling was performed using Vardict and Mutect2 where variants with VAF <5% were reported where clinically significant. These assays were performed by accredited pathology laboratories across Australia.

Cord blood and peripheral blood mononuclear cells from healthy donors

Umbilical cord blood was collected with written informed consent from scheduled cesarean section deliveries at the Women’s Health Unit, Lyell-McEwin Hospital (Adelaide, South Australia) between 12 November 2020 to 31 December 2023 with approval from the Women’s and Children’s Health Network Human Research Ethics Committee (HREC/20/WCHN/65; 2020/HRE01664). Peripheral blood buffy coat samples obtained with written informed consent was retrieved on 29 May 2023 and studies were approved by the Central Adelaide Local Health Network Human Research Ethics Committee (HREC/15/RAH/448) and conducted in accordance with the Declaration of Helsinki. Samples were processed by density gradient centrifugation using Lymphoprep TM (Stemcell Technologies, USA) to isolate mononuclear cells (MNCs).

Flow cytometry

Peripheral blood mononuclear cells (PB-MNCs) were collected at baseline from patients in the PREACH-M trial. 2–3 CBL mutant and 2–3 CBL wildtype CMML samples ( n = 4–6) were immunophenotyped by spectral flow cytometry (Cytek Aurora, USA). Control samples include cord blood mononuclear cells (CB-MNCs) ( n = 2) and PB-MNCs from healthy donors ( n = 1). All MNCs were incubated with Human TruStain FcX TM (BioLegend, USA) and True-Stain Monocyte Blocker TM (BioLegend, USA) prior to antibody staining. Antibody panel included mouse anti-human CD45 (HI30) BV421, CD14 (M5E2) PE-Cy7, CD16 (3G8) PE-Cy5, CD34 (8G12) APC, CD114 (LMM741) PE, CD116 (hGMCSFR-M1) PE, CD131 (1C1) PE; CD131 (3D7) BV421 and rat anti-human CD115 (9-4D2-1E4) PE. Viability was determined using Zombie Aqua TM Fixable Dye (BioLegend, USA). A list of catalogue numbers and clones of antibodies can be found in S1 Table .

Hotspot and protein structure analysis

Data from the Catalogue of Somatic Mutations in Cancer (COSMIC) were analyzed (last accessed on 15 February 2024). Analysis of CBL mutations were filtered as follows: Primary site hematopoietic and lymphoid, histology hematopoietic neoplasm; sub-histology CMML or JMML. 120 variants were found for CMML, 46 for JMML. Variants were stratified to include nonsense or missense substitutions, frameshift insertion or deletions, and in-frame insertions or deletions, with the exclusion of synonymous mutations, within the coding sequence. 17 coding region variants were detected in the PREACH-M cohort, making the total variants for CMML 137. The six most common mutations for CMML and JMML were identified. Protein structures were sourced from the Protein Data Bank (PDB) and analyzed using Maestro 13.8 (Schrodinger, USA) and UCSF ChimeraX (UCSF Resource for Biocomputing, Visualization and Information, USA). RMSD calculations were performed without rejection of any atoms during fit and distance was measured between alpha carbons of residue 227 and 371 of Cbl.

Statistical analysis

Values between unique samples were presented as mean ± standard error of margin (S.E.M.) or as mean ± standard deviation (S.D.) between technical replicates. For comparisons between groups, Student’s t -test or Mann-Whitney test was applied to analyze measurement (continuous) data and Fisher’s exact test for enumeration (categorical) data. All statistical analyses were performed using GraphPad Prism 10. P -values for Student’s t -tests were two-tailed, Mann-Whitney tests were one-tailed, and Fisher’s exact tests were two-tailed. P <0.05 was considered statistically significant.

CBL mutations are associated with increased marrow blasts, leukocytosis and splenomegaly, consistent with RAS pathway activation

Targeted NGS was performed on 24 de novo CMML baseline patient samples from the PREACH-M trial. Overall, RAS pathway ( KRAS , NRAS , PTPN11 , CBL ) mutations were detected in 18/24 (75%) patients with CBL mutations detected in 11/24 (46%) patients ( Tables 1 , 2 and S2 ). Consistent with previous findings where RAS pathway mutations were linked to the proliferative variant of CMML [ 54 , 55 ], patients with RAS pathway mutations had increased BM blast percentage (9.2 ± 1.1 vs . 5.3. ± 1.4%, P = 0.05), white cell count (WCC) (30.4 ± 6.0 vs . 13.8 ± 5.3×10 9 /L, P = 0.02), neutrophil absolute count (15.2 ± 3.3 vs . 7.2 ± 3.4×10 9 /L, P = 0.03), monocyte absolute count (7.8 ± 1.7 vs . 3.1 ± 1.1×10 9 /L, P = 0.03) and spleen length (15.2 ± 0.9 vs . 12.0 ± 0.5 cm, P = 0.04) compared to patients wildtype for any RAS pathway mutations ( Table 1 ). This finding led us to further stratify our cohort and indeed, patients with CBL mutations were also noted to have increased BM blast percentage (10.1 ± 1.5 vs . 5.3 ± 1.4%, P = 0.05), WCC (26.8 ± 5.9 vs . 13.8 ± 5.2×10 9 /L, P = 0.05), and spleen length (15.2 ± 1.1 vs . 12.0 ± 0.5 cm, P = 0.03) compared to CBL wildtype patients without RAS pathway mutations ( Table 2 and Fig 1A–1F ). Comparisons between CBL mutant/RAS pathway wildtype vs . RAS pathway mutant/ CBL wildtype cases revealed consistent trends ( S1 Fig ). 10/11 (90.9%; P = 0.03) patients with CBL variants presented with splenomegaly ( S3 Table ). This underscores the strong proliferative phenotype conferred by mutations in CBL in CMML. Importantly, 8/11 (73%) CBL variants were in cases classified as myeloproliferative-CMML (MP-CMML) based on WCC ( Fig 1G ), and 9/11 (82%) classified as CMML-1 or -2 based on BM blast percentage ( Fig 1H ) according to the 2016 WHO classification, linking CBL mutations not just to a proliferative phenotype but to more advanced stages of the disease, and therefore, to increased risk of progression to AML [ 50 ].

• PPT PowerPoint slide
• PNG larger image
• TIFF original image

( A-F ) Clinical characteristics of PREACH-M cohort at baseline, stratified based on the detection of CBL mutation. ( G ) MD-, MP-CMML classification based on WCC α ( H ) CMML-0, -1, -2 classifications based on BM blast percentage β . 2016 WHO Classification: α Based on WCC: MD-CMML WCC<13×10 9 /L, MP-CMML WCC>13×10 9 /L. β Based on BM blast %: CMML-0 PB <2%, BM <5%; CMML-1 PB 2–4%, BM 5–9%, CMML-2 PB>5%, BM 10–19%. Bars represent mean ± standard error of mean. Mann-Whitney test used to determine statistical significance, where P <0.05 was statistically significant. [BM bone marrow; WCC white cell count; CRP C-reactive protein; MD-CMML myelodysplastic-CMML; MP-CMML myeloproliferative-CMML, wt wildtype, mut mutant].

https://doi.org/10.1371/journal.pone.0310641.g001

https://doi.org/10.1371/journal.pone.0310641.t001

https://doi.org/10.1371/journal.pone.0310641.t002

CBL mutations co-occur frequently with TET2

Of the 24 patients in the study, 75% (18/24) were detected to have TET2 mutation, 58% (14/24) ASXL1 , 50% (12/24) SRSF2 and 46% (11/24) CBL ( Fig 2A ). Of the 11 patients with CBL mutation, 9 (82%) had co-occurring TET2 mutation. In 67% (6/9) of instances when these mutations are detected in the same patient, the difference in VAF magnitude between CBL and TET2 were ≤10% (MEL13, ADE02, ADE20, MEL05, MEL06, BRI07) ( Figs 2B and S2 ), indicating these mutations may co-occur within the same clone. On the contrary, mutations in CBL and other RAS pathway genes ( KRAS , NRAS , PTPN11 ) are not only infrequently found in the same patient (5/11; 45%), the VAF of the dominant CBL or RAS pathway mutant clone tend to be discordant, with VAF differences >10% (MEL13, BRI07, ADE17, ADE09), except in one case where CBL and PTPN11 VAF were ≤3% ( Figs 2B and S2 ).

( A ) Oncoplot for the PREACH-M cohort ( n = 24). Mutation groups are shown in rows with each individual patient represented by a column. The presence of a mutation is indicated by the red or blue colored bars. Age category of the patients indicated by the black and grey bars and sex of patients by the green and gold bars. ( B ) Number of CBL mutant cases where TET2 mutations ( n = 9) and other RAS pathway mutations ( n = 5) were detected, where variation in the VAF of CBL vs . TET2 or RAS pathway mutant clones were ≤10% (dark blue) or >10% (light blue) ( C ) Number of cases where more than one variant of CBL , NRAS , KRAS or PTPN11 mutation was detected. ( D ) Details of CBL variants detected in each patient with CBL mutation. [VAF variant allele frequency].

https://doi.org/10.1371/journal.pone.0310641.g002

Multiple CBL mutant subclones found in CMML

Strikingly, in 7/11 (64%) patients with CBL mutation, more than one CBL variant can be detected ( Fig 2C ). Of these, 2 patients had 3 variants while 5 patients had 2 variants ( Fig 2D ). In comparison, multiple subclones within a patient are more uncommon in KRAS (3/8; 38%), NRAS (2/5; 40%) and PTPN11 (0/3; 0%) mutant cases ( Fig 2C ).

CMML linked to high CD116 and CD131 in the progenitor subpopulation

As CMML is a disease characterized by upregulation of inflammatory cytokines [ 56 , 57 ] and expansion of pro-inflammatory granulocyte-macrophage-like progenitor cells and monocytes with enhanced cytokine receptor signaling [ 58 , 59 ], we analyzed the immunophenotype of primary patient samples ( n = 2–3 CBL mutant; n = 2–3 CBL wildtype) focusing on cytokine receptor expression, including granulocyte colony-stimulating factor receptor (G-CSFR, CD114), macrophage colony-stimulating factor receptor (M-CSFR, CD115) and the heterodimeric granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR) comprising the alpha subunit (GMRα, CD116) and the beta common subunit (βc, CD131), in the CD45 + MNCs, CD34 + hematopoietic stem and progenitor cells and CD14 + monocytes (gating strategy outlined in Fig 3A ). CMML patient samples had higher percentage of CD14 + cells compared to normal donors, consistent with expansion of monocytes and clinical presentation of the disease ( Fig 3B ).

( A ) Flow cytometry analysis of a representative CMML sample and healthy control stained for CD45, CD34, CD14 and CD16, and gating strategy used to define CD45 + mononuclear cells, CD34 + stem and progenitor cells and CD14 + monocytes. ( B ) Percentage of CD34 + progenitors and CD14 + monocytes in CMML samples ( n = 4) vs . healthy control ( n = 2). ( C ) Illustration of the cluster of differentiation (CD) markers where CD114 is a marker for G-CSFR, CD116 GMRα and CD131 βc. G-CSFR is homodimeric while GM-CSFR is heterodimeric receptor consisting of GMRα and βc. The expression of CD114, CD115, CD116 and CD131 in CMML samples ( n = 4–6) vs . control ( n = 2–3) in CD45 + , CD34 + and CD14 + subpopulations, expressed as percentage of positively stained cells ( D ) and MFI ( E ) compared to control (cord blood or peripheral blood mononuclear cells from healthy donors). Bars represent mean ± standard deviation in ( B ). Box and whiskers graphs were plotted with min and max in ( C ) and ( D ). Unpaired Student’s t -test between CMML vs . healthy control used to determine statistical significance, where P <0.05 was statistically significant. * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001. [MFI mean fluorescence intensity; G-CSFR granulocyte-colony stimulating factor receptor; GMRα granulocyte-macrophage colony stimulating factor receptor subunit α; βc beta common subunit].

https://doi.org/10.1371/journal.pone.0310641.g003

Our data revealed that the percentage of cells in CMML patient samples expressing CD116 (GMRα) was significantly higher in the total MNC population compared to control, and this was most pronounced in the CD34 + progenitor subpopulation (89.7 ± 1.6 vs . 50.3 ± 2.7%; P = 0.000003) ( Fig 3C and 3D ). In contrast, the percentage of CD116 expressing cells was similar between healthy and CMML-derived CD14 + monocytes ( Fig 3D ). Interestingly, the density of CD116 expression represented by mean fluorescence intensity (MFI) was upregulated in our CMML cohort vs . healthy controls (CD45 + 90.3 ± 4.6×10 3 vs . 30.0 ± 3.2×10 3 , P = 0.001; CD34 + 40.2 ± 7.4×10 3 vs . 13.9 ± 4.1×10 3 , P = 0.08; CD14 + 118.9×10 3 ± 8.1×10 3 vs . 77.4×10 3 ± 5.8×10 3 ; P = 0.03) ( Fig 3E ). We also noted an increase in the percentage of CD131 expression in the MNCs, particularly in the CD34 + progenitors in CMML compared to control (64.3 ± 3.8 vs . 32.1 ± 4.1%; P = 0.001), although not in terms of MFI ( Fig 3D and 3E ). We did not observe a difference in receptor expressions between CBL mutant and CBL wildtype CMML patient samples ( S3 Fig ).

We also noted increases in the percentage of CD114 + cells across all CMML cell populations (CD45 + P = 0.03; CD34 + P = 0.04; CD14 + P = 0.02), with no difference in the MFI ( Fig 3C and 3D ). In contrast, the percentage of CD115 + cells was notably lower in the CMML CD45 + MNCs ( P = 0.01) and CD14 + monocytes ( P = 0.005), with reductions in MFI seen in both CD34 + ( P = 0.005) and CD14 + ( P = 0.007) populations ( Fig 3D and 3E ).

CBL mutations are enriched in the RING domain in CMML compared to JMML

We then combined the new mutation data from our cohort with the publicly available data in COSMIC to assess which domains of Cbl were commonly perturbed. We found that CBL mutations in CMML and JMML are concentrated within the coding sequence of the LHR and RING domain of Cbl ( Fig 4A ). Furthermore, we noted that in CMML, mutations most frequently occur within the RING domain (amino acid residues 381–435) contrary to JMML, where mutations within the LHR (amino acid residues 353–380) are most common ( P <0.0001) ( Fig 4B ).

( A ) Table of CBL variants detected in our PREACH-M cohort combined with data sourced from COSMIC. Variants include nonsense or missense substitutions, frameshift and in-frame insertions or deletions within the coding sequence of CBL , filtered for all hematopoietic and lymphoid malignancies including CMML and JMML ( n = 549), and CMML only ( n = 137) or JMML only ( n = 46) ( B ) Contingency analysis of CBL mutation hotspots within the LHR and RING domain of Cbl in CMML and JMML ( C ) Heat map representation of all sites within the LHR (amino acid residues 353–380) and RING domain (amino acid residues 381–435) where mutations have been reported. Numbers within the figure and on the scale depict counts ( D ) Tertiary protein structure of native wildtype Cbl (PDB ID 2Y1M) in inactive, closed conformation. The TKBD is colored beige, LHR blue and RING domain red. Amino acid residues of the top 6 mutation hotspots are indicated in inset; Tyrosine 371 (Y371), Leucine 380 (L380), Cysteine 384 (C384), Cysteine 396 (C396), Cysteine 404 (C404) and Arginine 420 (R420). ( E ) X-ray structures of wildtype Cbl in unphosphorylated, inactive state and in closed conformation (PDB ID 2Y1M), wildtype Cbl in Y371 phosphorylated, active state and in open conformation (PDB ID 4A4C), mutant Cbl Y371E (PDB ID 5HKX) and mutant Cbl Y371F (PDB ID 5J3X). The TKBD is colored beige, LHR of wildtype blue, LHR of mutant cyan, RING domain of wildtype red, RING domain of mutant pink. RMSD values between various Cbl conformations are shown in table. Statistical analysis was performed using two-sided Fisher’s exact test, where P <0.05 was statistically significant. [TKBD tyrosine kinase binding domain; LHR linker helix region; RING RING domain; H+L all hematopoietic and lymphoid malignancies; WT wildtype; RMSD root mean square deviation (distanced-based measure of protein structure similarity)].

https://doi.org/10.1371/journal.pone.0310641.g004

The 6 most common CBL mutations in all hematopoietic and lymphoid malignancies occur in codons affecting amino acid residues 371, 380, 384, 396, 404, 420 ( Fig 4C and 4D ). In the PREACH-M cohort in particular, mutations in residue 384 were detected 3 patients, 371 in 2 patients, 380 in 2 patients, 404 and 420 in 1 patient, respectively. In CMML, missense substitutions cysteine 404 to tyrosine (C404Y) (13/137; 10%) and arginine 420 to glutamine (R420Q) (12/137; 9%) were most common, while in JMML, tyrosine 371 to histidine (Y371H) substitution was most common (21/46; 46%).

Mutations at residue 371 within the LHR can result in novel conformational change

Finally, we performed comparative structural alignments of available mutant Cbl structures resolved by X-ray diffraction publicly available via the Protein Data Bank (PDB). Comparison of the LHR of wildtype Cbl protein in the closed, inactive conformation (Y371 unphosphorylated) (PDB 2Y1M) [ 40 ] against the open, active conformation (Y371 phosphorylated) (PDB 4A4C) [ 40 ] ( Fig 4E ), revealed a root mean square deviation (RMSD) of 20.81Å, indicating that a significant conformational change takes place when Cbl becomes activated by phosphorylation. Further, we also compared PDB structures 2Y1M and 4A4C with structures comprising tyrosine 371 to glutamic acid (Y371E) (PDB 5HKX) and tyrosine 371 to phenylalanine (Y371F) (PDB 5J3X) [ 41 ] LHR mutations. Interestingly, we inferred that the Y371E mutant Cbl possesses an entirely different conformation to either closed, inactive or open, active wildtype Cbl (RMSD 9.49Å and 17.34Å, respectively). We noted that replacement of the polar, bulky tyrosine with the negatively charged glutamic acid (Y371E) resulted in perturbation of the LHR-TKBD interface and subsequent total displacement of the LHR and RING domain compared to both inactive and active wildtype Cbl ( Fig 4E ). In contrast, when tyrosine was replaced with a structurally similar residue phenylalanine (Y371F), the LHR-TKBD interface was unperturbed, and the mutant closely mimicked the native, inactive state of wildtype Cbl (RMSD 0.38Å) and not the active state (RMSD 20.88Å) ( Fig 4E ). Additional measurements relating to the structural differences between wildtype and mutant Cbl can be found in S4 Table .

Our data, obtained from patients enrolled in a prospective multicenter interventional study, highlight several clinical and molecular features of CBL mutants in CMML. Our results are generally consistent with previous studies that performed next generation sequencing in CMML patients but we note a higher frequency of CBL variants (11 of 24 patients, 46%) than others report (12.8%) [ 60 , 61 ], possibly due to strict trial eligibility criteria (higher white cell count or cytopenia). CBL variants were associated with a myeloproliferative phenotype, including higher white cell count and splenomegaly with many patients having increased blasts at diagnosis, similar to patients with other RAS pathway mutations. Notably, many patients harbored multiple CBL subclones (intrapatient molecular heterogeneity) that was not observed to the same extent for other RAS pathway mutations. This may be significant because subclonal abundance, especially a branched pattern of clonal evolution, is associated with a favorable outcome in AML [ 62 ].

We observed a strong overlap and clonal correlation between CBL mutations and TET2 mutation. We noted another study that has found a modest association of TET2 with CBL mutations (r<0.25; P<0.1) [ 60 ], despite the high frequency of TET2 mutation overall. A number of in vivo murine studies have highlighted a role for TET2 in suppressing innate immune signaling in monocytes, with TET2 mutant monocytes showing enhanced pro-inflammatory responses to stimuli such as lipopolysaccharide [ 63 , 64 ]. It is plausible that the CBL mutation serves to further amplify innate immune signaling by preventing ubiquitination and turnover of cytokine and Toll-like receptors but the exact cellular compartments within which this occurs is not defined. Thus, it is significant that a high percentage of CBL mutant CMML CD34 + progenitors express GMRα (CD116, 89.7%) and its partner subunit, βc (CD131, 64.3%), suggesting that a substantial proportion of CD34 + cells are primed to respond to the cytokine GM-CSF ( Fig 3C–3E ). Indeed, we know from previous studies that CMML display hypersensitivity to GM-CSF akin to JMML, especially in cases that have RAS pathway mutations [ 24 , 25 ]. In contrast, the percentage expression of the receptor for M-CSF (CD115) was decreased in CMML progenitors compared to controls, indicating this alternate monocyte cytokine is unlikely to be driving the disease. We also noted increased percentage of G-CSFR (CD114)-expressing CMML compared to controls, although not to the same extent as CD116, indicating CMML progenitors may also respond to G-CSF.

Previous studies with GM-CSF neutralizing antibody [ 25 ] and our own work with the GM-CSF E21R antagonist and successful engraftment of CMML patient samples in mice transgenic for human GM-CSF [ 24 ] provide strong evidence that GM-CSF is an essential growth factor for CMML in vitro and in vivo . Our findings that CD116 is upregulated in the CD34 + progenitors is interesting as it raises the question of the effect of GM-CSF on the leukemia-initiating cell population. This is in agreement with a recent study using single cell RNA-seq to map the differentiation trajectories of CD34 + progenitors in CMML primary patient samples [ 59 ] where the authors also showed the upregulation of CD116 in a cell cluster enriched for granulocyte/monocyte progenitor-like inflammatory hematopoietic stem and progenitor cells that may have self-renewal capacity in CMML patients with a monocyte-biased differentiation trajectory. Recently, we found that interleukin-3 (IL-3) receptor stoichiometry is a critical determinant in cell fate and IL-3 receptor overexpression in leukemia stem cells leads to biased activation of distinct transcriptional programs and signaling pathways to drive stemness programs vs . cell differentiation [ 65 ]. This propels us to hypothesize that although GM-CSF has been mostly associated with the proliferation and differentiation of hematopoietic progenitors and mature cells, it is possible that aside from the cytokine hypersensitivity previously shown [ 25 ], GM-CSF may also have unique signaling and effects on stem cell maintenance and function in CD116-overexpressing CD34 + progenitors for disease initiation and generation of the pro-inflammatory phenotype associated with this disease. Future studies should examine the kinetics of receptor turnover and phosphorylation peak and attenuation in this primary population, the signaling pathways involved and determine whether CD116 can be used to distinguish between CBL mutant leukemia vs . healthy stem cells. Indeed, the effects of anti-CD116 or anti-GM-CSF therapies (or in combination), on these populations warrant further investigation.

Cbl adopts a closed and open conformation dependent on Y371 phosphorylation to allow for the binding of ubiquitin-conjugating enzyme E2 [ 40 ]. Thus, it follows that the loss of this key tyrosine residue in position 371 within the LHR could have dramatic implications for the conformation and activity of Cbl. Tyrosine 371 is in a buried environment, where it forms a hydrogen bond with threonine 227 (T227) and makes several van der Waals interactions with residues in the hydrophobic pocket of the TKBD [ 36 , 40 ], playing a structural role in maintaining the integrity of the LHR-TKBD interface, and importantly, in keeping Cbl in a closed conformation, autoinhibited state [ 40 ]. When Y371 is phosphorylated, an open conformation is adopted, and autoinhibition is abolished leading to Cbl becoming a more active ligase [ 35 , 38 , 40 , 66 , 67 ]. Our analysis of the X-ray structures demonstrates that Cbl conformation is sensitive to the amino acid residue at position 371. Indeed, our inference following comparative structural alignments is that, depending on the nature of the substituting residue, mutation at position 371 (such as Y371E, PDB 5HKX) not only results in the impairment of phosphorylation-dependent activation, but can also yield an entirely novel conformation of Cbl, different from both the inactive (unphosphorylated, closed) and active (phosphorylated, open) conformations. With this conformational drift, the RING domain and E2 enzyme cannot be in sufficient proximity to the substrate binding site of the TKBD for effective ubiquitination. Thus, ubiquitination and degradation of activated RTKs would be predicted to occur less efficiently, resulting in sustained downstream signaling that may contribute to oncogenicity and disease progression. In contrast, mutations that do not perturb the LHR-TKBD interaction (such as Y371F, PDB 5J3X [ 41 ]) would mimic the conformation of native, wildtype Cbl, albeit no longer capable of increased catalytic efficiency due to loss of the phosphorylation site. This is consistent with early evidence that Cbl Y371 mutants can exist in different states of activity depending on the chemical nature of the amino acid substitution [ 41 ].

In CMML, CBL mutations were found in both the LHR and RING domains of the protein but with significant enrichment for mutations in the RING domain compared to JMML. The RING domain determines the specificity of Cbl E3 for its cognate E2 enzyme, recognizes lysines to be ubiquitinated and serves as a scaffold for optimal orientation for ubiquitin transfer between E2 and its substrate RTK [ 36 ] but the structure of Cbl RING domain mutations has not been determined. Elucidation of distinct mutant Cbl conformations is significant because new drug development strategies could employ proteolysis targeting chimera (PROTAC) technology for targeted protein degradation of mutant Cbl with conformations different from wildtype. Conversely, a new Cbl-b inhibitor C7683 currently in phase I clinical trials for advanced solid tumor malignancies, designed to keep wildtype Cbl-b locked in an inactive state [ 68 ], may partially mimic LHR Cbl mutations and thus should be used with caution in patients with clonal hematopoiesis, CMML or JMML.

A limitation of our study is the relatively small number of CBL mutant positive cases, reflecting the rarity of CMML. Nevertheless, certain clinical and molecular features are consistent across the cohort and are congruent with data available in COSMIC, implying that CBL mutant CMML may have a characteristic phenotype. Studies with larger cohorts are required to distinguish CBL mutation CMML from other RAS pathway mutations such as NRAS , KRAS and PTPN11 . To date, one study using serial VAF measurements did not show significant change in clone size for CMML patients, including CBL clones, treated with azacitidine alone suggesting epigenetic effects, rather than mutation-specific effects, were linked to therapeutic benefit [ 16 ]. In a phase I study, quizartinib inhibition of the receptor tyrosine kinase FLT3, a Cbl target for ubiquitination and internalization [ 69 ], did not appear to impact CMML with CBL mutations, suggesting phosphorylated FLT3 is not a critical substrate of Cbl in CMML [ 70 ]. Future work should examine the effect of CD116-targeted immunotherapy on the clonal dynamics of CBL RING domain vs . LHR mutants.

Supporting information

S1 table. list of antibodies for flow cytometry..

https://doi.org/10.1371/journal.pone.0310641.s001

S2 Table. Mutation status of PREACH-M cohort ( n = 24) with regards to CBL and other RAS pathway mutations ( KRAS , NRAS , PTPN11 ).

https://doi.org/10.1371/journal.pone.0310641.s002

S3 Table. Spleen craniocaudal length (cm) of CBL mutant CMML vs RAS pathway wildtype.

https://doi.org/10.1371/journal.pone.0310641.s003

S4 Table. Measurement of C α -C α distance between residue 227 and 371 δ from structural analysis of Cbl wildtype and mutant proteins.

https://doi.org/10.1371/journal.pone.0310641.s004

S1 Fig. Clinical characteristics of PREACH-M cohort at baseline, stratified according to CBL mutants and other RAS pathway ( KRAS , NRAS , PTPN11 ) mutants vs . wildtype.

https://doi.org/10.1371/journal.pone.0310641.s005

S2 Fig. VAF data of CBL mutant cases compared to TET2 and other RAS pathway genes.

https://doi.org/10.1371/journal.pone.0310641.s006

Cytokine receptor CD114, CD115, CD116 and CD131 expression of CBL mutant and wildtype compared to healthy control by (A) percentage positive cells and (B) mean fluorescence intensity (MFI).

https://doi.org/10.1371/journal.pone.0310641.s007

https://doi.org/10.1371/journal.pone.0310641.s008

Acknowledgments

The authors thank all patients and their families for donating specimens to research. The authors thank Verity Saunders and Kiralee Vuglar for isolation and cryopreservation of MNCs following blood collection from patients in the PREACH-M trial and Professor Gus Dekker and the Northern Adelaide Local Health Network for provision of fresh cord blood. Flow cytometry was performed at the Adelaide Health and BioMedical Precinct Cytometry Facility. Illustrations were created with BioRender.com.

• View Article
• PubMed/NCBI
• Google Scholar