Myeloblasts transition to megakaryoblastic immunophenotypes over time in some patients with myelodysplastic syndromes

Kiyoyuki Ogata; Yuto Mochimaru; Kazuma Sei; Naoya Kawahara; Mika Ogata; Yumi Yamamoto

doi:10.1371/journal.pone.0291662

Peer Review History

Original SubmissionApril 25, 2023
4 Jul 2023 Decision Letter - Daniel Thomas, Editor PONE-D-23-10670Myeloblasts transition to megakaryoblastic immunophenotypes over time in some patients with myelodysplastic syndromesPLOS ONE Dear Dr. Ogata, Thank you for submitting your manuscript to PLOS ONE. The paper is interesting and is of value to the field. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. Apologies for delay - we had difficulty obtaining timely responses from reviewers at this time. Please try and address to the suggestions by both reviewers. Please submit your revised manuscript by Aug 18 2023 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols. We look forward to receiving your revised manuscript. Kind regards, Daniel Thomas, MD Academic Editor PLOS ONE Journal requirements: When submitting your revision, we need you to address these additional requirements. 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. Additional Editor Comments: Ogata et al present real world flow cytometry data showing a reasonable proportion of high risk MDS patients express CD41 and can acquire CD41 over time, not due to platelet adherance/micromegakaryocytes. The findings are relevant and interesting given new data indicating megakaryocyte progenitors can arise directly from HSCs. A deficiency of the manuscript is the flow cytometry data is not linked closely with clinical data. Minor suggestions: 1] What proportion of CD41+ vs CD41- negative patients were actually t-MN, given the complex karyotypes noted. Should be discussed if no data. 2] Can we do a correlation plot of CD41 vs CD61 in the few patients tested and CD41 vs CD42b? Can we draw conclusions on what added benefit CD61 vs CD42b would have in the flow diagnostic panel? 3] Was CD110, TpoR, also expressed in any of the CD41+ blasts? 4] It would be desirable to present CD41+ (at any time) vs CD41- patients in a side by side table with P-values of significance - easy for the reader to see the differences 5] Early platelet recovery is a sign of azacitidine response - can we comment on the duration of azacitidine (+/- ven) for any of the CD41+ vs CD41- negative patients? [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Partly ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: N/A ****** 3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes ****** 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: Ogata and colleagues describe the expression of CD41 on CD34+ precursor cells in MDS or oligoblastic AML. This is not new, but extends on previous work from the same group. The main novelty is the report that acquisition of CD41 expression during azacytidine therapy (n=7) was associated with poor response or progression on therapy. Major comment The way that this study was undertaken introduces substantial bias. The patients were eligible for HMA therapy, so enriched for high risk. Serial bone marrow aspirates were performed only for poor responders and not systematically for all patients. This means that we simply don’t know whether CD41 expression might also arise in lower risk patients or those apparently responding to HMA. This should be acknowledged as a confounding factor. Minor comment Some of the references are incomplete (online prepublication details only) ****** 6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No ******** [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0291662.r001
Revision 1
11 Aug 2023 Author Response Dear Dr. Thomas, Thank you for reviewing and commenting on our manuscript titled “Myeloblasts transition to megakaryoblastic immunophenotypes over time in some patients with myelodysplastic syndromes.” We appreciate your valuable time and efforts. Here, I am resubmitting our manuscript, modified according to the comments from the editor and reviewer as well as the style requirements of PLOS ONE. The modified parts are highlighted in red in the revised manuscript. The original comments from the editor and reviewer and our responses to them are presented below in a point-by-point manner. We hope that the revised manuscript is now acceptable for publication in PLOS ONE. Thank you for your consideration. I look forward to hearing from you. Kiyoyuki Ogata, MD Responses to the editor The editor’s comment 1 What proportion of CD41+ vs CD41- negative patients were actually t-MN, given the complex karyotypes noted. Should be discussed if no data. Our response Ten patients in our cohort had therapy-related myeloid neoplasms (t-MN), which was unrelated to their CD41-positivity. In this revision, we showed these data in Table 1 and S2 Table. The editor’s comment 2 Can we do a correlation plot of CD41 vs CD61 in the few patients tested and CD41 vs CD42b? Can we draw conclusions on what added benefit CD61 vs CD42b would have in the flow diagnostic panel? Our response We examined the expression of CD41, CD42b, and CD61 on blasts in the same tube using flow cytometry; this was respectively done for five CD41+ patients, as described in the Methods and Table 2. In all five patients, the expression level of CD41 and CD61 on blasts showed a very strong correlation as shown in Figure 3 (Case 6). In the revised manuscript, we have added the plot of CD41 vs. CD61 for the other four cases (S2 Fig). Such a strong correlation between CD41 and CD61 expression is also observed in normal platelets (Ann Lab Med. 2014. 34(6): 471–474), as CD41 and CD61 molecules are known to form a gpIIb/IIIa complex. Further, in our five cases, addition of the CD61 vs. CD42b plot had no benefit in diagnosing patients with CD41+ MDS. However, because the number of cases was small, we avoided drawing a conclusion regarding the diagnostic value of adding CD61 (or the CD61 vs. CD42b plot); instead, we showed the CD41 vs. CD61 plots from all five cases as described above and added a comment on this issue (pages 14 and 18). As different laboratories are using different antibody panels in diagnostic flow cytometry, the data of CD61 expression in addition to those of CD41 and CD42b shown in this manuscript can be helpful to some laboratories. The editor’s comment 3 Was CD110, TpoR, also expressed in any of the CD41+ blasts? Our response This is an important issue from various viewpoints, including the positive or negative effect of TpoR agonists on patients with CD41+ MDS. Unfortunately, we have not yet examined TpoR expression in any of the cases. The editor’s comment 4 It would be desirable to present CD41+ (at any time) vs CD41- patients in a side by side table with P-values of significance - easy for the reader to see the differences. Our response In this revision, we created a new table (S2 Table), wherein basic characteristics were compared between CD41+ and CD41- patients in a side-by-side fashion with P-values. The editor’s comment 5 Early platelet recovery is a sign of azacitidine response - can we comment on the duration of azacitidine (+/- ven) for any of the CD41+ vs CD41- negative patients? Our response In this revision, we compared the number of azacitidine (+/- ven) cycles administered between the CD41+ and CD41- patients. More specifically, the number of cycles until the detection of CD41-positivity in former patients and that until the last flow cytometry analysis in latter patients were compared. However, we did not find a significant difference in the number (S2 Table). Responses to the reviewer Thank you very much for reviewing and commenting on our manuscript, which helped improve our manuscript. We highly appreciate your valuable time. The reviewer’s major comment The way that this study was undertaken introduces substantial bias. The patients were eligible for HMA therapy, so enriched for high risk. Serial bone marrow aspirates were performed only for poor responders and not systematically for all patients. This means that we simply don’t know whether CD41 expression might also arise in lower risk patients or those apparently responding to HMA. This should be acknowledged as a confounding factor. Our response We completely agree with this comment. We have acknowledged this important point in the discussion of the revised manuscript (page 19-20). The reviewer’s minor comment Some of the references are incomplete (online prepublication details only). Our response We have examined and corrected the references thoroughly. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0291662.r002
4 Sep 2023 Decision Letter - Daniel Thomas, Editor Myeloblasts transition to megakaryoblastic immunophenotypes over time in some patients with myelodysplastic syndromes PONE-D-23-10670R1 Dear Dr. Ogata, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Daniel Thomas, MD Academic Editor PLOS ONE Additional Editor Comments (optional): All comments have been satisfactorily addressed Reviewers' comments: NA https://doi.org/10.1371/journal.pone.0291662.r003
Formally Accepted
8 Sep 2023 Acceptance Letter - Daniel Thomas, Editor PONE-D-23-10670R1 Myeloblasts transition to megakaryoblastic immunophenotypes over time in some patients with myelodysplastic syndromes Dear Dr. Ogata: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Daniel Thomas Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0291662.r004

Open letter on the publication of peer review reports

PLOS recognizes the benefits of transparency in the peer review process. Therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. Reviewers remain anonymous, unless they choose to reveal their names.

We encourage other journals to join us in this initiative. We hope that our action inspires the community, including researchers, research funders, and research institutions, to recognize the benefits of published peer review reports for all parts of the research system.

Learn more at ASAPbio .