CTRP3 and serum triglycerides in children aged 7-10 years

Arsham Alamian; Jo-Ann Marrs; W. Andrew Clark; Kristy L. Thomas; Jonathan M. Peterson

doi:10.1371/journal.pone.0241813

Peer Review History

Original SubmissionJuly 21, 2020
15 Sep 2020 Decision Letter - Paolo Magni, Editor PONE-D-20-22639 CTRP3 and serum triglycerides in children aged 7-10 years PLOS ONE Dear Dr. Peterson, 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 address properly all reviewer's comments. Please submit your revised manuscript by October 5, 2020. 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: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols We look forward to receiving your revised manuscript. Kind regards, Paolo Magni 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 provide additional details regarding participant consent. In the ethics statement in the Methods and online submission information, please ensure that you have specified what type you obtained (for instance, written or verbal, and if verbal, how it was documented and witnessed). If your study included minors, state whether you obtained consent from parents or guardians. If the need for consent was waived by the ethics committee, please include this information. 3.Thank you for stating the following in the Funding Section of your manuscript: [This work was supported by grants from the Tennessee Board of Regents [Diversity Research Grant TBR (2014-2015)] and the National Institute of Diabetes and Digestive and Kidney Diseases [R15 DK114740-01A1]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.] We note that you have provided funding information that is not currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows: [The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.] Please include your amended statements within your cover letter; we will change the online submission form on your behalf. 4. Please include your tables as part of your main manuscript and remove the individual files. Please note that supplementary tables (should remain/ be uploaded) as separate "supporting information" files [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: 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: Summary: This study measured CTRP3 HMW and presumed MMW circulating levels from blood samples taken from pediatric donors. Key finding was that circulating triglycerides and VLDL levels negatively correlated with MMW CTRP3, while HMW CTRP3 did not show any statistically significant correlation with the measured parameters. Significance of study: The circulating levels of CTRP3 oligomeric complexes have previously been examined in an adult cohort with or without Type 2 Diabetes Mellitus. This is a similar study on a pediatric cohort. While the reported parameters are similar between the two studies, CTRP3 levels have not previously been examined in a pediatric demographic, and this study provides novel information relevant to the growing epidemic of childhood obesity and type 2 diabetes. General comments: The manuscript is clearly written except for some typographical errors throughout the manuscript and a few run-on sentences. Authors should carefully review the manuscript for typographical and grammatical errors. Appropriate statistical analyses have been performed on the data presented. Specific comments: 1) In Line 61, authors incorrectly state that adipose is the “largest endocrine organ by mass”. Muscle, the largest organ by mass, also secretes a vast array of bioactive molecules including myokines and cytokines. This statement should be corrected. 2) The authors use centrifugal separation to differentiate molecules above 300kDa from those under 300kDa, and conclude that CTRP3 measurements in these two fractions correspond to HMW and MMW CTRP3 populations. However, there is the possibility that the <300kDa fraction contains both LMW and MMW species, and the authors do not demonstrate experimentally that MMW CTRP3 is the only CTRP3 oligomeric species in this fraction. Although the authors cite a previous publication that reported no detectable LMW species in plasma, it should not be assumed that this is the case in a pediatric population. Authors should demonstrate that CTRP3 is not detected (by immunoblot or ELISA) in a <50kDa fraction. 3) Authors should include a schematic or table to summarize the direction of correlations between the measured parameters and HMW or MMW CTRP3 and total CTRP3. This will help the reader better visualize these patterns between the different CTRP3 oligomeric forms. 4) Typographical error in y-axis Fig 1b. 5) Data for HMW CTRP3 correlations with the measured parameters should be provided as a supplement or otherwise, as conclusions are drawn from these data. ****** 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. Attachments Attachment Submitted filename: PONE-D-20-22639_reviewer comments.docx https://doi.org/10.1371/journal.pone.0241813.r001
Revision 1
6 Oct 2020 Author Response Summary: This study measured CTRP3 HMW and presumed MMW circulating levels from blood samples taken from pediatric donors. Key finding was that circulating triglycerides and VLDL levels negatively correlated with MMW CTRP3, while HMW CTRP3 did not show any statistically significant correlation with the measured parameters. Significance: The circulating levels of CTRP3 oligomeric complexes have previously been examined in an adult cohort with or without Type 2 Diabetes Mellitus. This is a similar study on a pediatric cohort. While the reported parameters are similar between the two studies, CTRP3 levels have not previously been examined in a pediatric demographic, and this study provides novel information relevant to the growing epidemic of childhood obesity and type 2 diabetes. The manuscript is clearly written except for some typographical errors throughout the manuscript and a few run-on sentences. Authors should carefully review the manuscript for typographical and grammatical errors. Appropriate statistical analyses have been performed on the data presented. • We thank the reviewer for support of our manuscript and suggestions for improvement. We would like to apologize for typographical errors and oversights in the previous version and attest that all authors have carefully reviewed the revised version. Specific comments: 1) In Line 61, authors incorrectly state that adipose is the “largest endocrine organ by mass”. Muscle, the largest organ by mass, also secretes a vast array of bioactive molecules including myokines and cytokines. This statement should be corrected. • We have removed this statement as the reviewer is correct that in healthy weight persons muscle mass far exceeds adipose tissue. 2) The authors use centrifugal separation to differentiate molecules above 300kDa from those under 300kDa, and conclude that CTRP3 measurements in these two fractions correspond to HMW and MMW CTRP3 populations. However, there is the possibility that the <300kDa fraction contains both LMW and MMW species, and the authors do not demonstrate experimentally that MMW CTRP3 is the only CTRP3 oligomeric species in this fraction. Although the authors cite a previous publication that reported no detectable LMW species in plasma, it should not be assumed that this is the case in a pediatric population. Authors should demonstrate that CTRP3 is not detected (by immunoblot or ELISA) in a <50kDa fraction. • This is an excellent point by the reviewer. We performed additional experiments on these samples to attempt to detect CTRP3 in the <100 kDa fraction by immunoblot. We performed the assay multiple times with three different CTRP3 antibodies and we were unable to detect CTRP3 in the less than 100 kDa fraction (had we been able to detect CTRP3 <100 we would have also examined a <50 kDa fraction). We added a representative image as a supplemental file (Supplemental figure 1). 3) Authors should include a schematic or table to summarize the direction of correlations between the measured parameters and HMW or MMW CTRP3 and total CTRP3. This will help the reader better visualize these patterns between the different CTRP3 oligomeric forms. • We appreciate the reviewer’s suggestion and have added the requested information as 3 separate supplemental tables detailing the correlation coefficients between metabolic parameters and total CTRP3 (Table S1), HMW CTRP3 (Table S2), and MMW CTRP3 (Table S3). Attachments Attachment Submitted filename: PONE-D-20-22639_reviewer comments.docx https://doi.org/10.1371/journal.pone.0241813.r002
21 Oct 2020 Decision Letter - Paolo Magni, Editor CTRP3 and serum triglycerides in children aged 7-10 years PONE-D-20-22639R1 Dear Dr. Peterson, 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, Paolo Magni Academic Editor PLOS ONE Additional Editor Comments (optional): The comments have all been addressed. I recommend to follow this suggestion: It would be good practice to include a loading control for your western blot image in S1 to show that lack of detection of CTRP3 in the <100Kda fractions is not due to total protein not passing through the filtration process. Transferrin (77Kd) is a good loading control. 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: (No Response) ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: (No Response) ****** 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: (No Response) ****** 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: Thank you for addressing the comments. Additionally, it would be good practice to include a loading control for your western blot image in S1 to show that lack of detection of CTRP3 in the <100Kda fractions is not due to total protein not passing through the filtration process. Transferrin (77Kd) is a good loading control. ****** 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.0241813.r003
Formally Accepted
13 Nov 2020 Acceptance Letter - Paolo Magni, Editor PONE-D-20-22639R1 CTRP3 and serum triglycerides in children aged 7-10 years Dear Dr. Peterson: 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. Paolo Magni Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0241813.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 .