Severity of non-alcoholic steatohepatitis is not linked to testosterone concentration in patients with type 2 diabetes

Kristin Alexandra Dayton; Fernando Bril; Diana Barb; Jinping Lai; Srilaxmi Kalavalapalli; Kenneth Cusi

doi:10.1371/journal.pone.0251449

Peer Review History

Original SubmissionJanuary 19, 2021
26 Mar 2021 Decision Letter - Massimo Federici, Editor PONE-D-21-02015 Severity of Non-alcoholic Steatohepatitis is Not Linked to Testosterone Concentration in Patients with Type 2 Diabetes PLOS ONE Dear Dr. Cusi, 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 Apr 29 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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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, Massimo Federici, M.D. Academic Editor PLOS ONE Additional Editor Comments: comments from reviewer 1 I read with interest the manuscript of Dayton et al regarding the influence of liver disease on testosterone in males withT2DM. In this cross-sectional study of 175 males with T2DM, NAFLD was linked to lower total testosterone in patients with T2DM, but likely given a common soil of insulin resistance/obesity and not from the severity of liver necroinflammation or fibrosis. The manuscript is interesting and well written. I have some points: - In figure 1, could you perform also a test to evaluate intra-group differences? - In figure 4, could you perform also a test to evaluate intra-group differences for panel A and D? - In Figure 3, were the variables logaritimically transformed? Based on Table 1, they seems not normally distribuited - In page 12 the authors wrote: “In order to assess whether the association between total plasma testosterone and intrahepatic triglyceride content was independent of other metabolic factors, we performed a multiple linear regression analysis. Intrahepatic triglyceride content was independently associated with total testosterone levels (ß=-4.8, p=0.004). Body mass index (ß=-9.6, p<0.001) and HbA1c (ß=-19.8, p=0.02) were also independently associated with total testosterone in the multiple linear regression”. Please, could you report a table with these data? 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. Thank you for stating the following in the Acknowledgments Section of your manuscript: "Grant support for this study was provided by the Burroughs Wellcome Fund (K. C.), the American Diabetes Association (1-08-CR-08 [K. C.]), and a VA Merit Award (1 I01 CX000167-01 [K. C.])." 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: "American Diabetes Association (1-08-CR-08 [K. C.]) https://www.diabetes.org/ VA Merit Award (1 I01 CX000167-01 [K. C.]), https://www.research.va.gov/funding/ 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. 3. Please include captions for your Supporting Information files at the end of your manuscript, and update any in-text citations to match accordingly. Please see our Supporting Information guidelines for more information: http://journals.plos.org/plosone/s/supporting-information. [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: I read with interest the manuscript of Dayton et al regarding the influence of liver disease on testosterone in males withT2DM. In this cross-sectional study of 175 males with T2DM, NAFLD was linked to lower total testosterone in patients with T2DM, but likely given a common soil of insulin resistance/obesity and not from the severity of liver necroinflammation or fibrosis. The manuscript is interesting and well written. I have some points: - In figure 1, could you perform also a test to evaluate intra-group differences? - In figure 4, could you perform also a test to evaluate intra-group differences for panel A and D? - In Figure 3, were the variables logaritimically transformed? Based on Table 1, they seems not normally distribuited - In page 12 the authors wrote: “In order to assess whether the association between total plasma testosterone and intrahepatic triglyceride content was independent of other metabolic factors, we performed a multiple linear regression analysis. Intrahepatic triglyceride content was independently associated with total testosterone levels (ß=-4.8, p=0.004). Body mass index (ß=-9.6, p<0.001) and HbA1c (ß=-19.8, p=0.02) were also independently associated with total testosterone in the multiple linear regression”. Please, could you report a table with 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. 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Revision 1
9 Apr 2021 Author Response (letter has been attached earlier) Massimo Federici, M.D. Academic Editor PLOS ONE March 26th, 2021 RE: PLOS ONE Resubmission of manuscript PONE-D-21-02015. Dear Dr. Federici, We are resubmitting for publication in PLOS One our revised version of the manuscript titled entitled “Severity of NASH is Not Linked to Testosterone Concentration in Patients with Type 2 Diabetes” by Dayton et al. We appreciate the kind words of support by the editors and reviewers, as well as their excellent suggestions to enhance our manuscript. We include below a point-by-point response to the issues raised. We hope that the referees and editorial board will find this improved version to meet the high standards of your prestigious Journal. Best regards, Kenneth Cusi, M.D., F.A.C.P., F.A.C.E. Professor of Medicine Response to referee Referee #1: I read with interest the manuscript of Dayton et al regarding the influence of liver disease on testosterone in males withT2DM. In this cross-sectional study of 175 males with T2DM, NAFLD was linked to lower total testosterone in patients with T2DM, but likely given a common soil of insulin resistance/obesity and not from the severity of liver necroinflammation or fibrosis. Thank you for your support. The manuscript is interesting and well written. I have some points: Q1. In figure 1, could you perform also a test to evaluate intra-group differences? After adjustment for multiple comparisons, between-group comparisons in Figure 1 are: Group 1 vs. 2: p = 0.19 Group 2 vs. 3: p = 0.99 Group 1 vs. 3: p = 0.13 (The p we used was a p for trend so it makes sense that comparisons between groups are NS) Data included in the legend (page 10). Q2. In figure 4, could you perform also a test to evaluate intra-group differences for panel A and D? Fig 4 A. After adjustment for multiple comparisons, between-group comparisons are: Steatosis grade 1 vs. 2: p = 0.003 Steatosis grade 2 vs. 3: p = 0.99 Steatosis grade 1 vs. 3: p = 0.37 Fig4 D. After adjustment for multiple comparisons, between-stage groups comparisons are: Stage 0 vs. 1: 0.56 Stage 1 vs. 2: 0.99 Stage 0 vs. 2: 0.65 Stage 0 vs. 3: 0.99 Stage 2 vs. 3: 0.99 Data included in the legend (page 12). Q3. In Figure 3, were the variables logarithmically transformed? Based on Table 1, they seem not normally distributed. We appreciate this important question from the reviewer. In Figure 3 variables were not logarithmically transformed because only liver fat and HOMA were not normally distributed based on Shapiro-Wilk normality tests. Testing these variables after logarithmic transformation, produced similar correlations to those reported in Fig. 3: Total testosterone and liver fat: -0.28, p=0.002. Total testosterone and HOMA: -0.30, p<0.001. Due to the fact that the relationships were similar, for simplicity of interpretation for the readers, we decided to present the data without log transformation. Q4. In page 12 the authors wrote: “In order to assess whether the association between total plasma testosterone and intrahepatic triglyceride content was independent of other metabolic factors, we performed a multiple linear regression analysis. Intrahepatic triglyceride content was independently associated with total testosterone levels (ß=-4.8, p=0.004). Body mass index (ß=-9.6, p<0.001) and HbA1c (ß=-19.8, p=0.02) were also independently associated with total testosterone in the multiple linear regression”. Please, could you report a table with these data? This is a great suggestion and will add to the scientific merit of the paper. The table (below) was created and added to the manuscript as well. Coefficient p 95% confidence interval Liver fat content -4.8 0.004 -7.9 -1.6 HbA1c -19.8 0.024 -37.0 -2.7 BMI -9.6 <0.001 -13.8 -5.3 ALT 0.8 0.016 0.2 1.5 Table 2: Results of multiple linear regression analysis for variables associated with total testosterone. HDL, TG, Age, FPG, AST were also tested but were not independently associated with total testosterone. Attachments Attachment Submitted filename: Response_to_reviewers.docx https://doi.org/10.1371/journal.pone.0251449.r002
27 Apr 2021 Decision Letter - Massimo Federici, Editor Severity of Non-alcoholic Steatohepatitis is not Linked to Testosterone Concentration in Patients with Type 2 Diabetes PONE-D-21-02015R1 Dear Dr. Cusi, 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, Massimo Federici, M.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: Thank you very much for addressing my points. The maunuscript has been improved. No further comment. ****** 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.0251449.r003
Formally Accepted
21 May 2021 Acceptance Letter - Massimo Federici, Editor PONE-D-21-02015R1 Severity of Non-alcoholic Steatohepatitis is not Linked to Testosterone Concentration in Patients with Type 2 Diabetes Dear Dr. Cusi: 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. Massimo Federici Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0251449.r004

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