Up-regulation of plasma miRNA-21 and miRNA-422a in postmenopausal osteoporosis

Neda Mohammadisima; Azizeh Farshbaf-khalili; Alireza Ostadrahimi

doi:10.1371/journal.pone.0287458

Peer Review History

Original SubmissionDecember 12, 2022
10 Apr 2023 Decision Letter - Gary S. Stein, Editor PONE-D-22-34068Up-regulation of plasma miRNA-21 and miRNA-422a in postmenopausal osteoporosisPLOS ONE Dear Dr. Farshbaf-Khalili, 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 May 25 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. 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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 research is to verify the association between plasma miRNA-21-5p and miRNA-422a with osteoporosis in postmenopausal women. Certain questions have yet to be clarified. 1. Though miRNAs as biomarkers of postmenopausal osteoporosis have been studied intensively, certain miRNAs expressed in plasma at different stage of postmenopausal osteoporosis are still not clear. In current study, plasma miRNA-21-5p and miRNA-422a were chosen to verify their association with osteoporosis in postmenopausal women. Conclusion: “these miRNAs' expression levels recommend as helpful markers for screening women at risk of osteoporosis.” Which is more helpful or both? Method and Materials Participants: “They were referred to the Bone Densitometry center. Based on densitometric tests, 142 normal- 85 BMD, 109 osteoporosis, and 194 osteopenia women were identified.” It is suggested that the expression of miRNA-21-5p and miRNA-422a should be detected in some women qualified the inclusion criteria of 194 osteopenia women. Thus, the candidate miRNAs as biomarkers of postmenopausal patients in the early stage may be further explored through the comparison of osteoporosis, osteopenia, and normal groups, and maybe contribute to predict the occurrence of postmenopausal osteoporosis. 2. Potential target genes of miRNA-21-5p and miRNA-422a can be predicted through bioinformatic analysis and subsequently verified. 3. The primer sequences used for qRT-PCR should be listed as attached table. 4. Please standardize the statistical symbol p as lowercase and italic type. In general, the current research seems to be not deep enough. Reviewer #2: In this manuscript, the authors have investigated the association between plasma miRNA-21-5p and miRNA-422a with osteoporosis in postmenopausal women. By evaluating 65 osteoporotic and 61 normal cases of women aged 50-65, it has been concluded that miRNA-21 and miRNA-422 are significantly upregulated in osteoporotic women compared to normal. The results further reveal that there are inverse correlations between the levels of miRNA-21 and miRNA-422a and bone mineral density of spine and femoral neck in osteoporotic samples. The role of miRNA-422a and miRNA21 have been investigated in postmenopausal osteoporosis. For example, miRNA 422a has been studied in osteoclast precursor cells as a biomarker and miRNA 21 has been shown to regulate bone turn over markers. This investigation using plasma miRNAs further confirms that miRNAs 422a and 21 could be explored as diagnostic markers in postmenopausal osteoporosis. Minor comment: 1.Although, the results are quite convincing and interesting, the sample size (65 osteoporotic and 61 normal) is too small. 2. The manuscript has few typographical errors that need to be fixed. ****** 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". 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Revision 1
26 Apr 2023 Author Response Title: Up-regulation of plasma miRNA-21 and miRNA-422a in postmenopausal osteoporosis Dear Editor-in-Chief, I hope you are well and thank a lot for your attention to our submitted manuscript. Following are our responses to the Editor in Chief’s and reviewers’ comments. I hope that our corrections will be satisfactory for the respected editor. Kind regards PONE-D-22-34068R1 We've checked your submission and before we can proceed, we need you to address the following issues: 1. Thank you for stating the following financial disclosure: [No-The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.] At this time, please address the following queries: a) Please clarify the sources of funding (financial or material support) for your study. List the grants or organizations that supported your study, including funding received from your institution. Response: The Vice-chancellor for Research and Technology, Tabriz University of Medical Sciences financially funded the original research (grant no: 61494). b) State what role the funders took in the study. If the funders had no role in your study, please state: “The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.” Response: It was written. The funders had no role in your study, please state: “The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. c) If any authors received a salary from any of your funders, please state which authors and which funders. Response: No any authors received a salary from the funder. d) If you did not receive any funding for this study, please state: “The authors received no specific funding for this work.” Response: The authors received no specific funding for this work. 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Response: Please let us upload the data as Supporting Information files Kind regards, Reviewer #1: 1. Though miRNAs as biomarkers of postmenopausal osteoporosis have been studied intensively, certain miRNAs expressed in plasma at different stage of postmenopausal osteoporosis are still not clear. In current study, plasma miRNA-21-5p and miRNA-422a were chosen to verify their association with osteoporosis in postmenopausal women. Conclusion: “these miRNAs' expression levels recommend as helpful markers for screening women at risk of osteoporosis.” Which is more helpful or both? Response: Thanks a lot for your attention to our submitted manuscript. In our study, the expression of both miRNAs in postmenopausal women with osteoporosis is higher than normal. miRNA-21 and miRNA-422a had good and excellent diagnostic accuracy, respectively. Diagnostic accuracy of BTMs was explained in abstract and main text. Figure 3 was added in this regard. Method and Materials Participants: “They were referred to the Bone Densitometry center. Based on densitometric tests, 142 normal- 85 BMD, 109 osteoporosis, and 194 osteopenia women were identified.” It is suggested that the expression of miRNA-21-5p and miRNA-422a should be detected in some women qualified the inclusion criteria of 194 osteopenia women. Thus, the candidate miRNAs as biomarkers of postmenopausal patients in the early stage may be further explored through the comparison of osteoporosis, osteopenia, and normal groups, and maybe contribute to predict the occurrence of postmenopausal osteoporosis. Response: Thank a lot for your suggestion. 2. Potential target genes of miRNA-21-5p and miRNA-422a can be predicted through bioinformatic analysis and subsequently verified. Response: Thank a lot for your attention to our submitted manuscript. This was added to the suggestions at the end of discussion. 3. The primer sequences used for qRT-PCR should be listed as attached table. Response: The primer sequences used for qRT-PCR were listed in table 1. 4. Please standardize the statistical symbol p as lowercase and italic type. Response: statistical symbol p changed to lowercase and italic type. Reviewer #2: In this manuscript, the authors have investigated the association between plasma miRNA-21-5p and miRNA-422a with osteoporosis in postmenopausal women. By evaluating 65 osteoporotic and 61 normal cases of women aged 50-65, it has been concluded that miRNA-21 and miRNA-422 are significantly upregulated in osteoporotic women compared to normal. The results further reveal that there are inverse correlations between the levels of miRNA-21 and miRNA-422a and bone mineral density of spine and femoral neck in osteoporotic samples. The role of miRNA-422a and miRNA21 have been investigated in postmenopausal osteoporosis. For example, miRNA 422a has been studied in osteoclast precursor cells as a biomarker and miRNA 21 has been shown to regulate bone turn over markers. This investigation using plasma miRNAs further confirms that miRNAs 422a and 21 could be explored as diagnostic markers in postmenopausal osteoporosis. Minor comment: 1. Although, the results are quite convincing and interesting, the sample size (65 osteoporotic and 61 normal) is too small. Response: Thank you. We referred to it in limitations. 2. The manuscript has few typographical errors that need to be fixed. Response: Typographical errors were solved as possible. Thank a lot for your attention to our submitted manuscript. Regards Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0287458.r002
6 Jun 2023 Decision Letter - Gary S. Stein, Editor Up-regulation of plasma miRNA-21 and miRNA-422a in postmenopausal osteoporosis PONE-D-22-34068R1 Dear Dr. Farshbaf-Khalili, 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, Gary S. Stein Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0287458.r003
Formally Accepted
13 Jun 2023 Acceptance Letter - Gary S. Stein, Editor PONE-D-22-34068R1 Up-regulation of plasma miRNA-21 and miRNA-422a in postmenopausal osteoporosis Dear Dr. Farshbaf-Khalili: 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. Gary S. Stein Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0287458.r004

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