Evaluation of a marker independent isolation method for circulating tumor cells in esophageal adenocarcinoma

Annouck Philippron; Lieven Depypere; Steffi Oeyen; Bram De Laere; Charlotte Vandeputte; Philippe Nafteux; Katleen De Preter; Piet Pattyn

doi:10.1371/journal.pone.0251052

Peer Review History

Original SubmissionJanuary 4, 2021
17 Mar 2021 Decision Letter - Dominique Heymann, Editor PONE-D-21-00260 EVALUATION OF A MARKER INDEPENDENT ISOLATION METHOD FOR CIRCULATING TUMOR CELLS IN ESOPHAGEAL ADENOCARCINOMA PLOS ONE Dear Dr. Philippron, Thank you for submitting your manuscript to PLOS ONE. 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. Please submit your revised manuscript by 10th May 2021. 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. 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Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols We look forward to receiving your revised manuscript. Kind regards, Dominique Heymann, Ph.D. 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 [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: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes ****** 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: The manuscript submitted by Philippron et al, entitled “EVALUATION OF A MARKER INDEPENDENT ISOLATION METHOD FOR CIRCULATING TUMOR CELLS IN ESOPHAGEAL ADENOCARCINOMA” focus on the optimization of a marker-indipendent CTC enrichment which is Parsortix in particular in esophageal cancer patients. Authors aim attention at first to compare the gold standard Cellsearch and Parsortix technologies to enrich putative CTCs (OE33 and OE19 cell lines). Authors found that CellSearch had a higher capture rate instead of Parsortix, but they chose to perform CTCs enrichment on esophageal cancer patients only by the Parsortix platform, Authors should explain better the reason for this choice. Authors performed a morphology analysis of putative CTCs though DEPArray technology, but it is not clear why Authors did not use morphology analysis for patient samples. This is a matter of concern since authors often highlighted the potential usefulness of CTC analysis as well as molecular characterization; but they did not perform any type of this kind of analysis. More specifically Lines: 25 “the molecular characterization of circulating tumor cells (CTCs)...” and Line 39 “aiming towards downstream single-cell molecular characterization” authors stated these on the abstract objective and methods sections, but they did not perform this analysis; this kind of assumption should be addressed only in the discussion section this point is misleading. Overall this manuscript shed a tiny light on the CTCs analysis in blood of esophageal cancer patients, but authors should pay attention to do not overstate their findings, I referred especially with DEPArray analysis which was not performed on blood patients and molecular analysis not performed as well. ****** 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.0251052.r001
Revision 1
29 Mar 2021 Author Response Comments Reviewer #1: 1. Reviewer: Authors aim attention at first to compare the gold standard Cellsearch and Parsortix technologies to enrich putative CTCs (OE33 and OE19 cell lines). Authors found that CellSearch had a higher capture rate instead of Parsortix, but they chose to perform CTCs enrichment on esophageal cancer patients only by the Parsortix platform, Authors should explain better the reason for this choice. Authors: We chose the Parsortix platform to be evaluated, benchmarked against the Cellsearch platform. After completing the comparison between the two platforms using esophageal cell lines, we continued with Parsortix only on patient samples. We did not take extra blood samples for analysis on Cellsearch because this was not the goal set forward in this study. 2. Reviewer: Authors performed a morphology analysis of putative CTCs though DEPArray technology, but it is not clear why Authors did not use morphology analysis for patient samples. This is a matter of concern since authors often highlighted the potential usefulness of CTC analysis as well as molecular characterization; but they did not perform any type of this kind of analysis. More specifically Lines: 25 “the molecular characterization of circulating tumor cells (CTCs)...” and Line 39 “aiming towards downstream single-cell molecular characterization” authors stated these on the abstract objective and methods sections, but they did not perform this analysis; this kind of assumption should be addressed only in the discussion section this point is misleading. Authors: We realise this can be misleading. We did not perform the same analysis with DEPArray on the patient samples because the number of CTC’s per blood tube from an esophageal cancer patient in currative setting is simply too few. Future aspects in this study are that a higher CTC prevalence can be found in metastatic patients which can make CTC analysis- including molecular characterization- possible. We will adapt all reference to ‘molecular characterization’ in the manuscript to avoid misinterpretation and add this aspect to future perspectives at the end in the section ‘Discussion’. 3. Reviewer: Overall this manuscript shed a tiny light on the CTCs analysis in blood of esophageal cancer patients, but authors should pay attention to do not overstate their findings, I referred especially with DEPArray analysis which was not performed on blood patients and molecular analysis not performed as well, descriptive manuscript, carefull to not overstate findings, we carefully reviewed this manuscript Authors: We agree with the reviewer and will revise this descriptive manuscript to not overstate our findings. However, we were carfull to write a ‘descriptive’ manuscript and to not formulate statements because of the more explorative study with low number of patient samples. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0251052.r002
20 Apr 2021 Decision Letter - Dominique Heymann, Editor EVALUATION OF A MARKER INDEPENDENT ISOLATION METHOD FOR CIRCULATING TUMOR CELLS IN ESOPHAGEAL ADENOCARCINOMA PONE-D-21-00260R1 Dear Dr. Philippron, 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, Dominique Heymann, Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation. Reviewer #1: All comments have been addressed ******** 2. 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: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes ****** 4. 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 ****** 5. 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 ****** 6. 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: (No Response) ****** 7. 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 https://doi.org/10.1371/journal.pone.0251052.r003
Formally Accepted
29 Apr 2021 Acceptance Letter - Dominique Heymann, Editor PONE-D-21-00260R1 EVALUATION OF A MARKER INDEPENDENT ISOLATION METHOD FOR CIRCULATING TUMOR CELLS IN ESOPHAGEAL ADENOCARCINOMA Dear Dr. Philippron: 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 Pr. Dominique Heymann Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0251052.r004

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