Mesenchymal stem cell-neural progenitors are enriched in cell signaling molecules implicated in their therapeutic effect in multiple sclerosis

Violaine K. Harris; Jaina Wollowitz; Jacelyn Greenwald; Alyssa L. Carlson; Saud A. Sadiq

doi:10.1371/journal.pone.0290069

Peer Review History

Original SubmissionFebruary 10, 2023
24 Apr 2023 Decision Letter - Mária A. Deli, Editor PONE-D-23-03957Mesenchymal stem cell-neural progenitors are enriched in cell signaling molecules implicated in their therapeutic effect in multiple sclerosis.PLOS ONE Dear Dr. Harris, 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 all the points raised during the review process. ==============================Two experts have evaluated the manuscript and further amendments are suggested. The gene expression data are well analyzed and and the addition of protein data for selected genes well supports the main conclusions of the study. No further experiments are needed, but in the Discussion the limitations of the present study should be addressed.============================== Please submit your revised manuscript by Jun 08 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'. 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If consent was waived for your study, please include this information in your statement as well. [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 Reviewer #2: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: 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 Reviewer #2: 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 Reviewer #2: 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 use of MSC cells in regenerative medicine is a very important and promising field. The discovery of the mechanisms behind the positive effects is particularly important when the cell products are put into clinical trials. From this point of view, the work can provide useful and interesting results in the exploration of the mechanism of MSCs and MSC-NPs. Although the number of samples is low in this study (as the authors properly point it out), I consider it important that the samples came from the already completed clinical trial, so no further invasive intervention was necessary. The data seem to be well processed, and the conclusions are moderate. I would like to make only a few comments, mainly concerning the discussion part as follows: - in Figure 1A, although I agree that there is no difference between the MS and control groups, it is not clear why the authors say that the samples are clustered into one group? Rather, it cannot be interpreted as clustering or interpret what it means in this case if they are not clustered together. The differences between MS donors are addressed again in the discussion "Furthermore, examination of the >100 MS donor-derived MSCs manufactured for autologous administration into clinical trial subjects (cliniclatrials.gov ID NCT01933802, NCT03355365 and NCT03822858) has not revealed any correlation between donor age or disease duration and population doubling time of MSCs (data not shown).” That's a very strong claim without presenting the data, especially as this is a controversial issue in the field. I recommend rephrasing the sentence or providing supporting data. - Due to the spheroid shape, it is completely understandable that the expression of extracellular matrix genes changes in MSC-NPs, but what relevance can this have in vivo conditions? This should also be addressed in the discussion. Reviewer #2: The paper was described correctly and clearly. Unfortunately, with 2 control donors, it is difficult to draw conclusions. However, the main experimental gaps relate to further research. The evaluation of gene expression seems to be only the beginning. Subsequently, the level of proteins and the secretory properties of the cells should be assessed. Changes in gene expression in differentiated cells are obvious. The question is whether they have an impact on the secretion and potential therapeutic properties of the cells. If the authors carry out further experiments, then the paper will be very interesting. ****** 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 Reviewer #2: 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.0290069.r001
Revision 1
17 May 2023 Author Response Reviewer #1: The use of MSC cells in regenerative medicine is a very important and promising field. The discovery of the mechanisms behind the positive effects is particularly important when the cell products are put into clinical trials. From this point of view, the work can provide useful and interesting results in the exploration of the mechanism of MSCs and MSC-NPs. Although the number of samples is low in this study (as the authors properly point it out), I consider it important that the samples came from the already completed clinical trial, so no further invasive intervention was necessary. The data seem to be well processed, and the conclusions are moderate. I would like to make only a few comments, mainly concerning the discussion part as follows: - in Figure 1A, although I agree that there is no difference between the MS and control groups, it is not clear why the authors say that the samples are clustered into one group? Rather, it cannot be interpreted as clustering or interpret what it means in this case if they are not clustered together. Author response: The reviewer is correct in pointing out that the term ‘clustering’ was inappropriately used in the sentence. The Results section on page 10 of the manuscript was revised to say the following “PCA analysis (Fig 1) showed minimal variance between MSCs across groups.” The differences between MS donors are addressed again in the discussion "Furthermore, examination of the >100 MS donor-derived MSCs manufactured for autologous administration into clinical trial subjects (cliniclatrials.gov ID NCT01933802, NCT03355365 and NCT03822858) has not revealed any correlation between donor age or disease duration and population doubling time of MSCs (data not shown).” That's a very strong claim without presenting the data, especially as this is a controversial issue in the field. I recommend rephrasing the sentence or providing supporting data. Author response: The above-mentioned sentence has been removed from the revised manuscript, as the point is sufficiently made without it. - Due to the spheroid shape, it is completely understandable that the expression of extracellular matrix genes changes in MSC-NPs, but what relevance can this have in vivo conditions? This should also be addressed in the discussion. Author response: The following paragraph and references were added to the Discussion page 22: “ECM proteins are abundantly expressed in MSCs where they are well-known to play a key role in MSC-mediated tissue regeneration by restoring tissue structure, directing cell migration, promoting angiogenesis and stimulating nerve growth [14]. Although the upregulation of ECM proteins in MSC-NPs is likely related to their spheroid morphology, there is evidence that they also play a role in enhanced cell survival and regenerative function [22]. TIMP-1, for example, has been shown to promote oligodendrocyte differentiation and enhance CNS myelination, suggesting that MSC-NP-derived TIMP-1 may play a therapeutic role in the context of MS [23, 24]. Similarly, osteopontin, which is significantly upregulated in MSC-NPs, has been shown to promote oligodendrocyte proliferation and differentiation suggesting that it is beneficial to remyelination [25]. While further validation is required, the upregulation of ECM proteins in MSC-NPs points to a potential therapeutic mechanism by which MSC-NPs may modify neuroinflammation and remyelination in MS.” Reviewer #2: The paper was described correctly and clearly. Unfortunately, with 2 control donors, it is difficult to draw conclusions. However, the main experimental gaps relate to further research. The evaluation of gene expression seems to be only the beginning. Subsequently, the level of proteins and the secretory properties of the cells should be assessed. Changes in gene expression in differentiated cells are obvious. The question is whether they have an impact on the secretion and potential therapeutic properties of the cells. If the authors carry out further experiments, then the paper will be very interesting. Author response: We have confirmed increased protein expression/secretion for several of the genes described in manuscript, including PEDF and HGF in Figure 4, and TIMP-1 and Osteopontin in Figure 6. Additional validation of protein secretion and the contribution of these proteins to the therapeutic properties of MSC-NPs, while outside the scope of this particular study, is an important future direction of this research. We revised the last sentence in the manuscript discussion to highlight this point. “Whether or not these gene expression differences translate into increased secretion of specific proteins that impact the potency or therapeutic efficacy of MSC-NP-based cell therapy remains to be determined.” Attachments Attachment Submitted filename: AUTHOR RESPONSE LETTER.docx https://doi.org/10.1371/journal.pone.0290069.r002
2 Aug 2023 Decision Letter - Mária A. Deli, Editor Mesenchymal stem cell-neural progenitors are enriched in cell signaling molecules implicated in their therapeutic effect in multiple sclerosis. PONE-D-23-03957R1 Dear Dr. Harris, 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, Mária A. Deli, M.D., 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: In my opinion, the manuscript improved during the review process and the authors gave satisfactory answers to all my questions. ****** 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.0290069.r003
Formally Accepted
4 Aug 2023 Acceptance Letter - Mária A. Deli, Editor PONE-D-23-03957R1 Mesenchymal stem cell-neural progenitors are enriched in cell signaling molecules implicated in their therapeutic effect in multiple sclerosis. Dear Dr. Harris: 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 Prof. Mária A. Deli Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0290069.r004

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