Associations of FGF21 and GDF15 with mitochondrial dysfunction in children living with perinatally-acquired HIV: A cross-sectional evaluation of pediatric AIDS clinical trials group 219/219C

Greg S. Gojanovich; Denise L. Jacobson; Carly Broadwell; Brad Karalius; Brian Kirmse; Mitchell E. Geffner; Jennifer Jao; Russell B. Van Dyke; Elizabeth J. McFarland; Margarita Silio; Marilyn Crain; Mariana Gerschenson; for the Pediatric HIV/AIDS Cohort Study

doi:10.1371/journal.pone.0261563

Peer Review History

Original SubmissionJuly 21, 2021
24 Sep 2021 Decision Letter - Qigui Yu, Editor PONE-D-21-22547Associations of FGF21 and GDF15 with mitochondrial dysfunction in children living with perinatally-acquired HIV: a cross-sectional evaluation of Pediatric AIDS Clinical Trials Group 219/219CPLOS ONE Dear Dr. Gerschenson, 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 Nov 08 2021 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. 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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 If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. 2. You indicated that you had ethical approval for your study. In your Methods section, please ensure you have also stated whether you obtained consent from parents or guardians of the minors included in the study or whether the research ethics committee or IRB specifically waived the need for their consent. Additional Editor Comments (if provided): [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 Reviewer #3: Partly ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: 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 Reviewer #3: No ****** 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 Reviewer #3: 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: This study looked at the relationship between two biomarkers, FGF21 and GDF15, and their relationship to Mitochondrial Disease in CPHIV. Metabolic and mitochondrial pathology are hallmarks of HIV infection due to its’ propensity for manipulating host metabolism via viral proteins. Mitokines, such as those in this study, are produced as a result of mitochondrial stress and can therefore indication HIV driven mitochondrial dysfunction. The authors were able to find increased trends of both mitokines with increasing severity of disease, however, they did not find correlations with viral load (which should be highlighted more in the study). Furthermore, their usage of “significant correlation” is not correct as an R of greater than 0.5 would have been required. All correlations found in this study (other than control FGF21 v. GDF15) were minor to moderate. Samples from individuals with confirmed mitochondrial disease would have been helpful. The addition of mitochondrial dysfunction measurements in each of the groups would also have been helpful. Despite this, the paper does sufficiently show evidence for their claims. Reviewer #2: It’s a well-designed study, the statistical methods used are appropriate. I have no concerns about this study except one question: the mitochondrial diseases in most cases are inherited, do you have data in related to the CPHIV parents’ mitochondrial conditions and tried to analyze the association of mitokines between parents and children? Reviewer #3: A clinical trial was conducted which aimed to assess the relationship between FGF21 and GDF15 levels in children with mitochondrial disease (MD) and HIV. A matched pairs analysis was conducted for those with MDC of 4. Unmatched cases with MDC of 3 were also assessed. FGF21 and GDF15 were highest in MDC4, moderate in MDC3 and lowest in controls. Minor revisions: 1- At each mention of the Wilcoxon test, specify the exact type. 2- The estimate for a correlation coefficient is represented by r instead of rho. 3- The p-value associated with a correlation is a test of the null hypothesis: correlation equal to zero; however, the absolute magnitude of the coefficient indicates the strength of the linear relationship between two variables. In general, the strength or correlation coefficient is the more important statistic to focus on. Below is a table for interpreting correlation coefficients: Coefficient (absolute value) Interpretation 0.90 - 1.0 Very Strong 0.70 - 0.89 Strong 0.40 - 0.69 Moderate 0.10 - 0.39 Weak less than 0.10 Negligible correlation 4- Indicate the date range subjects were enrolled in the study. 5- Cite the statistical software used for the analysis. 6- State and justify the study’s target sample size with a pre-study statistical power calculation. The power calculation should include: (1) the estimated outcomes in each group; (2) the α (type I) error level; (3) the statistical power (or the β (type II) error level); (4) the target sample size and (5) for continuous outcomes, the standard deviation of the measurements. ****** 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 Reviewer #3: 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.0261563.r001
Revision 1
12 Nov 2021 Author Response Response to Reviewer Comments “Associations of FGF21 and GDF15 with mitochondrial dysfunction in children living with perinatally-acquired HIV: a cross-sectional evaluation of Pediatric AIDS Clinical Trials Group 219/219C " We would like to thank the editor and reviewers for their time and expertise with reviewing our manuscript. We agree that the suggested revisions make for a more robust and accurate manuscript, and have adapted it accordingly. We appreciate the reviewers stating that our manuscript is ‘a well-designed study, the statistical methods used are appropriate’ and that ‘the paper does sufficiently show evidence for their claims.’ Please see responses to specific reviewer remarks below in italics, and track changes have been used to highlight alterations within the manuscript, while mention of specific lines refers to this track-changed version. An unmarked, updated version without track changes is also supplied. Editor’s Comments to the Authors 1. 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. Authors’ Response: We have found no retraction notices for any articles cited, however corrections were published to amend minor concerns in Montero et al. and Tsygankova et al., which did not result in meaningful differences to conclusions drawn upon herein. Additionally, a formatting error for reference notation within the manuscript was corrected on line 218. 2. You indicated that you had ethical approval for your study. In your Methods section, please ensure you have also stated whether you obtained consent from parents or guardians of the minors included in the study or whether the research ethics committee or IRB specifically waived the need for their consent. Authors’ Response: We have amended our wording to indicate that we obtained the participant’s guardians consent on line 121. Reviewers’ Comments to the Authors: Reviewer #1: This study looked at the relationship between two biomarkers, FGF21 and GDF15, and their relationship to Mitochondrial Disease in CPHIV. Metabolic and mitochondrial pathology are hallmarks of HIV infection due to its’ propensity for manipulating host metabolism via viral proteins. Mitokines, such as those in this study, are produced as a result of mitochondrial stress and can therefore indication HIV driven mitochondrial dysfunction. The authors were able to find increased trends of both mitokines with increasing severity of disease, however, they did not find correlations with viral load (which should be highlighted more in the study). Furthermore, their usage of “significant correlation” is not correct as an R of greater than 0.5 would have been required. All correlations found in this study (other than control FGF21 v. GDF15) were minor to moderate. Samples from individuals with confirmed mitochondrial disease would have been helpful. The addition of mitochondrial dysfunction measurements in each of the groups would also have been helpful. Despite this, the paper does sufficiently show evidence for their claims. Authors’ Response: We agree with the reviewer’s concern regarding significance statements relating to the correlations and have adjusted phrasing accordingly, with a specific change found on line 430 in the discussion. We also agree that having samples from individuals with confirmed mitochondrial disease would have bolstered our data here, however this proposed sample collection was outside of the scope of our study, and the subjects within our study were not consented for histological/biochemical analyses that would have permitted clinical confirmation. Reviewer #2: It’s a well-designed study, the statistical methods used are appropriate. I have no concerns about this study except one question: the mitochondrial diseases in most cases are inherited, do you have data in related to the CPHIV parents’ mitochondrial conditions and tried to analyze the association of mitokines between parents and children? Authors’ Response: The authors completely agree that this line of study is warranted and, in fact, have already proposed such studies be performed in mother/child pairs, but are awaiting funding for this proposal. Reviewer #3: A clinical trial was conducted which aimed to assess the relationship between FGF21 and GDF15 levels in children with mitochondrial disease (MD) and HIV. A matched pairs analysis was conducted for those with MDC of 4. Unmatched cases with MDC of 3 were also assessed. FGF21 and GDF15 were highest in MDC4, moderate in MDC3 and lowest in controls. Minor revisions: 1- At each mention of the Wilcoxon test, specify the exact type. 2- The estimate for a correlation coefficient is represented by r instead of rho. 3- The p-value associated with a correlation is a test of the null hypothesis: correlation equal to zero; however, the absolute magnitude of the coefficient indicates the strength of the linear relationship between two variables. In general, the strength or correlation coefficient is the more important statistic to focus on. Below is a table for interpreting correlation coefficients: Coefficient (absolute value) Interpretation 0.90 - 1.0 Very Strong 0.70 - 0.89 Strong 0.40 - 0.69 Moderate 0.10 - 0.39 Weak less than 0.10 Negligible correlation 4- Indicate the date range subjects were enrolled in the study. 5- Cite the statistical software used for the analysis. 6- State and justify the study’s target sample size with a pre-study statistical power calculation. The power calculation should include: (1) the estimated outcomes in each group; (2) the α (type I) error level; (3) the statistical power (or the β (type II) error level); (4) the target sample size and (5) for continuous outcomes, the standard deviation of the measurements. Authors’ Response: 1. We agree and have adjusted accordingly throughout the manuscript. 2. We have changed all instances of rho to r. 3. We have changed wording selection to align with these suggestions throughout the manuscript. 4. We have moved this manuscript section from the results to the methods so as to be more similar to commonly used formatting. 5. Thank you for catching this oversight; this can now be found on line 255. 6. A statement regarding power calculations can now be found starting on line 236. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0261563.r002
6 Dec 2021 Decision Letter - Qigui Yu, Editor Associations of FGF21 and GDF15 with mitochondrial dysfunction in children living with perinatally-acquired HIV: a cross-sectional evaluation of Pediatric AIDS Clinical Trials Group 219/219C PONE-D-21-22547R1 Dear Dr. Gerschenson,, 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, Qigui Yu, M.D./Ph.D Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0261563.r003
Formally Accepted
10 Dec 2021 Acceptance Letter - Qigui Yu, Editor PONE-D-21-22547R1 Associations of FGF21 and GDF15 with mitochondrial dysfunction in children living with perinatally-acquired HIV: a cross-sectional evaluation of Pediatric AIDS Clinical Trials Group 219/219C Dear Dr. Gerschenson: 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. Qigui Yu Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0261563.r004

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