Mechanisms involved in suppression of osteoclast supportive activity by transforming growth factor-β1 via the ubiquitin-proteasome system

Momoko Inoue; Yoshie Nagai-Yoshioka; Ryota Yamasaki; Tatsuo Kawamoto; Tatsuji Nishihara; Wataru Ariyoshi

doi:10.1371/journal.pone.0262612

Peer Review History

Original SubmissionAugust 18, 2021
2 Nov 2021 Decision Letter - Xing-Ming Shi, Editor PONE-D-21-26829Mechanisms involved in suppression of osteoclast supportive activity by transforming growth factor-β1 via the ubiquitin-proteasome systemPLOS ONE Dear Dr. Ariyoshi, 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 discuss the in vitro and in vivo effects of TGF-beta1 on osteoclasts and emphasize what new information this study contributes to the field. Please submit your revised manuscript by Dec 16 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. 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: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols. We look forward to receiving your revised manuscript. Kind regards, Xing-Ming Shi, Ph.D 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. PLOS ONE now requires that authors provide the original uncropped and unadjusted images underlying all blot or gel results reported in a submission’s figures or Supporting Information files. This policy and the journal’s other requirements for blot/gel reporting and figure preparation are described in detail at https://journals.plos.org/plosone/s/figures#loc-blot-and-gel-reporting-requirements and https://journals.plos.org/plosone/s/figures#loc-preparing-figures-from-image-files. When you submit your revised manuscript, please ensure that your figures adhere fully to these guidelines and provide the original underlying images for all blot or gel data reported in your submission. See the following link for instructions on providing the original image data: https://journals.plos.org/plosone/s/figures#loc-original-images-for-blots-and-gels. In your cover letter, please note whether your blot/gel image data are in Supporting Information or posted at a public data repository, provide the repository URL if relevant, and provide specific details as to which raw blot/gel images, if any, are not available. Email us at plosone@plos.org if you have any questions. 3. Please ensure that you refer to Figure 2 in your text as, if accepted, production will need this reference to link the reader to the figure. [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: No 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: No 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: Comment 1) Effects of TGF-beta1 in vivo and in vitro. In this study, the authors demonstrated TGF-beta 1 down-regulated RANKL expression in vitro. TGF-beta1 is well known to exert suppressive effects in vitro, and to exert promotive effects in vivo. Actually, the many manuscript has been published already regarding TGF-beta 1 derived from bone matrix promoted osteoclastgenesis. Therefore, this study demonstrated one-sided effects of TGF-beta 1 and effects of TGF-beta 1 is already known to the potential readers. The authros should revise the manuscript extensively so that the potential readers can find new findings, and recognize this study does not contain one-sided effects. Comment 2) Stromal cells or mesenchymal stem cells ? Comment 3) Many sentences involving unclear words and meanings Reviewer #2: This manuscript described the important role of the transforming growth factor-β1 (TGF-β1) on the osteoclast differentiation. In this paper, TGF-β1 downregulates RANKL expression and the osteoclast-supporting activity of osteoblasts/stromal cells through the degradation of the RXR-α protein mediated by ubiquitin proteasome system. It should be discussed in detail whether the findings clarified in this paper can explain the mechanism of action of TGF-β1 on osteoclast differentiation by bone resorption factors other than vitamin D, PTH and PGE2. ****** 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: Yes:** Nobuyuki Udagawa [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: Comment.docx https://doi.org/10.1371/journal.pone.0262612.r001
Revision 1
16 Nov 2021 Author Response Comment 1) Effects of TGF-β1 in vivo and in vitro. In this study, the authors demonstrated TGF-β1 down-regulated RANKL expression in vitro. TGF-β1 is well known to exert suppressive effects in vitro, and to exert promotive effects in vivo. Actually, the many manuscripts have been published already regarding TGF-β1 derived from bone matrix promoted osteoclastgenesis. Therefore, this study demonstrated one-sided effects of TGF-beta 1 and effects of TGF-β1 is already known to the potential readers. The authors should revise the manuscript extensively so that the potential readers can find new findings, and recognize this study does not contain one-sided effects. We appreciate your important suggestions. Several studies demonstrated that the role of TGF-β1 on bone remodeling process. However, the reported effects of TGF-β1 are controversial. Since many of these studies are focused on the direct effects of TGF-β1 on osteoclast progenitors, we investigated the effects of TGF-β1 on RANKL generation in stromal cells. Moreover, the role of TGF-β1 as a coupling factor of bone remodeling process should be elucidated. However, we did not mention about it. We edited the Discussion session in the revised manuscript with some references. Comment 2) Stromal cells or mesenchymal stem cells? In general, stromal cells involves all the non-epithelial cells including fibroblast, lipocytes, lymphcytes, muscular cells, and neuron cells. Only in the field of regeneration, a stromal cell is referred to as a mesenchymal stem cell (MSC). To avoid misunderstanding of the potential readers, the stromal cells in the current study should be revised as mesenchymal stem cells. We appreciate your helpful suggestions. ST2 cell is a cloned stromal cell line from mouse bone marrow. This cell line has the ability to differentiate into osteoblast-like cells and shows high expression of RANKL and low expression of OPG when treated with 1,25(OH)2D3 and dexamethasone. However, we did not mention about it. So, we added this information in the Materials and methods section of revised manuscript. Comment 3) Many sentences involving unclear words and meanings. There are many sentences involving unclear words and meanings. These sentences should be revised before submission. The following are the examples. We appreciate your helpful comments. We carefully checked and revised the sentences that were pointed out. Example 1) The current study investigated the effect of TGF-β1 on receptor activator of nuclear factor kappa-B ligand (RANKL) expression, induced by 1α,25(OH)2D3 (D3) and dexamethasone (Dex) in stromal cells. This is very complex. RANKL expression in stromal cell Thank you for your helpful comments. We edited the abstract section of revised manuscript. Example 2) Stromal cells support osteoclastogenesis.; Please explain the meaning of "support osteoclastogenesis” in this study. Is this mean "induce osteoclastogenesis"? What is the difference? Thank you for your suggestions. Previous studies have been revealed the direct effect of TGF-β1 on the osteoclast differentiation from osteoclast progenitor cells. In this study, we showed that TGF-β1 indirectly inhibits the expression of RANKL, an accelerator of osteoclast differentiation in stromal cells. To emphasize these differences, we used the term “support osteoclastogeneis” in the place of “induce osteoclastogenesis”. Example 3) ST2 cells in the abstract section. Please explain what ST2 cells mean in the abstract section. As noted by the reviewer’s comment, we added the information of ST2 cells in the abstract section. Example 4) Transforming growth factor-β1 (TGF-β1) is a coupling factor vital for bone remodeling. What means "vital bone remodeling"? As noted by the reviewer’s comment, we edited the abstract section of revised manuscript. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0262612.r002
30 Dec 2021 Decision Letter - Xing-Ming Shi, Editor Mechanisms involved in suppression of osteoclast supportive activity by transforming growth factor-β1 via the ubiquitin-proteasome system PONE-D-21-26829R1 Dear Dr. Ariyoshi, 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, Xing-Ming Shi, Ph.D Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0262612.r003
Formally Accepted
14 Feb 2022 Acceptance Letter - Xing-Ming Shi, Editor PONE-D-21-26829R1 Mechanisms involved in suppression of osteoclast supportive activity by transforming growth factor-β1 via the ubiquitin-proteasome system Dear Dr. Ariyoshi: 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 Xing-Ming Shi Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0262612.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 .