Galectin-9 as a biomarker for disease activity in systemic lupus erythematosus

Naoki Matsuoka; Yuya Fujita; Jumpei Temmoku; Makiko Yashiro Furuya; Tomoyuki Asano; Shuzo Sato; Haruki Matsumoto; Hiroko Kobayashi; Hiroshi Watanabe; Eiji Suzuki; Hideko Kozuru; Hiroshi Yastuhashi; Kiyoshi Migita

doi:10.1371/journal.pone.0227069

Peer Review History

Original SubmissionSeptember 29, 2019
15 Oct 2019 Decision Letter - Masataka Kuwana, Editor PONE-D-19-27302 Galectin-9 as a biomarker for disease activity in systemic lupus erythematosus PLOS ONE Dear Dr. Migita, 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. Both reviewers found some interests in this article, but there are a number of points that require improvement, or oven amendment. Especially, comments made by reviewer #1 is critical. Submission of revised version requires thorough responses to all comments made by reviewers, and massive rewriting the manuscript. We would appreciate receiving your revised manuscript by Nov 29 2019 11:59PM. When you are 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. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. To enhance the reproducibility of your results, we recommend that if applicable you deposit your laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols 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). This letter should be uploaded as separate file and labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. This file should be uploaded as separate file and labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. This file should be uploaded as separate file and labeled 'Manuscript'. Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out. We look forward to receiving your revised manuscript. Kind regards, Masataka Kuwana, MD, PhD 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 http://www.journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and http://www.journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. Thank you for stating the following in your Competing Interests section: 'The study was supported by the Practical Research Project for Rare / Intractable Diseases from Japan Agency for Medical Research and Development, AMED.' Please complete your Competing Interests on the online submission form to state any Competing Interests. If you have no competing interests, please state "The authors have declared that no competing interests exist.", as detailed online in our guide for authors at http://journals.plos.org/plosone/s/submit-now This information should be included in your cover letter; we will change the online submission form on your behalf. Please know it is PLOS ONE policy for corresponding authors to declare, on behalf of all authors, all potential competing interests for the purposes of transparency. PLOS defines a competing interest as anything that interferes with, or could reasonably be perceived as interfering with, the full and objective presentation, peer review, editorial decision-making, or publication of research or non-research articles submitted to one of the journals. Competing interests can be financial or non-financial, professional, or personal. Competing interests can arise in relationship to an organization or another person. Please follow this link to our website for more details on competing interests: http://journals.plos.org/plosone/s/competing-interests 3. 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 4. 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. 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: Partly Reviewer #2: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: No 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: First of all, although the authors claimed that Galectin-9 (Gal-9) could be a serologic marker of disease activity and organ damage in systemic lupus erythematosus (SLE) patients, a single biomarker is rarely associated with SLE disease activity and damage concomitantly because damage in SLE patients is irreversible change in organs usually due to longstanding disease activity. Thus, that would rather raise suspicion over the accuracy of the SLEDAI-2K scores and SLICC damage index in the present study. Alternatively, the true association between Gal-9 and SLE. This issue should be clarified in the manuscript. In addition, although the authors reported that Gal-9 is elevated in CSF from patients with NPSLE and claimed that the present study showed the clinical significance of CSF Gal-9, most of the previously reported candidate CSF biomarkers for NPSLE were elevated exclusively in CSF but not in serum. Thus, that would rather raise suspicion over the significance of Gal-9 as a biomarker for SLE. This issue should be clarified in the manuscript, too. In the abstract, “damage scores” should be written as the SLICC damage index. In the abstract, values such as “16.6 ng/ml [3.6-59.5]” should be explained that they were medians and 5–95th centiles. In the abstract, although the authors wrote that levels of Gal-9 were also significantly higher in the CSF from patients with recent-onset neuropsychiatric SLE (NPSLE) than in those from non-SLE controls, the representative values and statistics should be mentioned. In the Methods section, original papers should be cited for SLEDAI-2K and SLICC damage index instead of review articles. In the Submission Guidelines of this journal, it is stated that all research involving human participants must have been conducted according to the principles expressed in the Declaration of Helsinki. This should probably be mentioned in the manuscript. In the Methods section, it was stated that 7 patients with NPSLE were included outside of the enrolled 58 SLE paints in whom serum samples were not collected. These 7 NPLSE patients should be included in the study population. Moreover, it is difficult to understand why these 7 NPLSE patients were excluded from the serum analyses. In the Methods section, it was stated that results were non-normally distributed and were compared by the Mann-Whitney U test. Then why correlations between continuous variables were analyzed by the Pearson correlation test?. In the Results section, p values were reported for correlation analyses. However, the correlation is independent of sample size, whereas the p value is affected by sample size. Thus, correlation coefficients and their confidence intervals are more important than p values. In the Discussion section, the first several sentences (from “SLE is a systemic autoimmune condition” to “SLE disease activity and organ involvements” should be discussed in the Introduction section. In the Discussion section of the manuscript, the authors claimed that Gal-9 is a more powerful discriminator between SLE patients with organ damage compared with CXCL10 and M2BPGi. If the authors believe this, ROC curve analyses should be performed and the sensitivity and specificity for the thresholds should be demonstrated. From the dot plots where many values are in the same ranges, it does not seem that serum levels of Gal-9 are able to discriminate SLE patients with and without organ damage, though. Throughout the manuscripts, there are grammatical errors. Reviewer #2: Authors demonstrated that galectin-9 is a serological marker of disease activity and organ damage in SLE patients. The new correlation between galectin-9 titer and organ damage is different from the previous paper (ref # 7). 1)Discuss why galectin-9 increases with organ damage from the perspective of cells that produce galectin-9. 2)Galectin-9 is also a biomarker in other autoimmune diseases, such as primary sjogren syndrome (ARD in press, 2018-214651) and juvenile dermatomyositis (A & R 71 (8): 1377-90, 2019) and correlates with disease activity in each disease. It is not a marker specific to SLE. Please discuss from this point of view. ****** 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". If this link does not appear, there are no attachment files to be viewed.] 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 us at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0227069.r001
Revision 1
25 Nov 2019 Author Response Reviewer #1: First of all, although the authors claimed that Galectin-9 (Gal-9) could be a serologic marker of disease activity and organ damage in systemic lupus erythematosus (SLE) patients, a single biomarker is rarely associated with SLE disease activity and damage concomitantly because damage in SLE patients is irreversible change in organs usually due to longstanding disease activity. Thus, that would rather raise suspicion over theaccuracy of the SLEDAI-2K scores and SLICC damage index in the present study. Alternatively, the true association between Gal-9 and SLE. This issue should be clarified in the manuscript. We appreciate your critical comments. The SDI scores irreversible organ damage that occurred since the onset of SLE regardless of cause. The British Isles lupus assessment Group (BILAG) is used to evaluate the specific manifestations and to be useful tool in monitoring disease activity in SLE patients. According to your critical comments, we evaluated the disease activity and SLE-related organ involvement using this BILAG sore. We presented these new data and discuss this important point in the revised manuscript. In addition, although the authors reported that Gal-9 is elevated in CSF from patients with NPSLE and claimed that the present study showed the clinical significance of CSF Gal-9, most of the previously reported candidate CSF biomarkers for NPSLE were elevated exclusively in CSF but not in serum. Thus, that would rather raise suspicion over the significance of Gal-9 as a biomarker for SLE. This issue should be clarified in the manuscript, too. We appreciate your critical comments. According to your important comments, we compared CSF Gal-9 with serum Gal-9 in patients with NPSLE and controls. As pointed out in your comment, our data suggested that CSF Gal-9 levels seems to be elevated regardless of the values of serum Gal-9 levels in a subset of NPSLE patients. We added these data in the revised manuscript. In the abstract, “damage scores” should be written as the SLICC damage index. In the abstract, values such as “16.6 ng/ml [3.6-59.5]” should be explained that they were medians and 5–95th centiles. According to your precise comments, we corrected and described the range of the presented data as interquartile range (IQR). In the abstract, although the authors wrote that levels of Gal-9 were also significantly higher in the CSF from patients with recent-onset neuropsychiatric SLE (NPSLE) than in those from non-SLE controls, the representative values and statistics should be mentioned. According to your precise comments, we added these data in the Abstract. In the Methods section, original papers should be cited for SLEDAI-2Kand SLICC damage index instead of review articles. We appreciate your critical comments. We cited the proper reference forSLEDAI-2K and SLICC damage index in the revised manuscript. In the Submission Guidelines of this journal, it is stated that all research involving human participants must have been conducted according to the principles expressed in the Declaration of Helsinki. This should probably be mentioned in the manuscript. We appreciate your critical comments. We mentioned this important issues including ethnical approval in the revised manuscript. In the Methods section, it was stated that 7 patients with NPSLE were included outside of the enrolled 58 SLE paints in whom serum samples were not collected. These 7 NPLSE patients should be included in the study population. Moreover, it is difficult to understand why these 7 NPLSE patients were excluded from the serum analyses. We appreciate your critical comments. Due to the small number of the paired samples, (CSF plus sera), we could not analyze the CSF plus sera in a sufficient number of patients. In the Methods section, it was stated thatresults were non-normally distributed and were compared by the Mann-Whitney U test. Then why correlations between continuous variables were analyzed by the Pearson correlation test?. In the Results section, p values were reported for correlation analyses. However, the correlation is independent of sample size, whereas the p value is affected by sample size. Thus, correlation coefficients and their confidence intervals are more important than p values. We appreciate your critical comments. According to your precise comments, we evaluated the corrections between Gal-9 and clinical parameters using Spearman’s correction test. We also calculated the confidence intervals. We corrected these descriptions and added these data in revised manuscript. In the Discussion section, the first several sentences (from “SLE is a systemic autoimmune condition” to “SLE disease activity and organ involvements” should be discussed in the Introduction section. We appreciate your precise comments. According to your comments, we revised these description and discuss this important point in the revised manuscript. In the Discussion section of the manuscript, the authors claimed that Gal-9 is a more powerful discriminator between SLE patients with organ damage compared with CXCL10 and M2BPGi. If the authors believe this, ROC curve analyses should be performed and the sensitivity andspecificity for the thresholds should be demonstrated. From the dot plots where many values are in the same ranges, it does not seem that serum levels of Gal-9 are able to discriminate SLE patients with and without organ damage, though. We appreciate your critical comments. According to your comments, it is difficult to discriminate SLE patients with or without organ damage using this single biomarker (Galectin-9), therefore, we deleted these descriptions in the revised manuscript. Throughout the manuscripts, there are grammatical errors. Reviewer #2: Authors demonstrated that galectin-9 is a serological marker of disease activity and organ damage in SLE patients. The newcorrelation between galectin-9 titer and organ damage is different from the previous paper (ref # 7). 1)Discuss why galectin-9 increases with organ damage from the perspective of cells that produce galectin-9. We appreciate your critical comments. According to your comments, we discussed this important point in the revised manuscript. 2)Galectin-9 is also a biomarker in other autoimmune diseases, such as primary sjogren syndrome (ARD in press, 2018-214651) and juvenile dermatomyositis (A & R 71 (8): 1377-90, 2019) and correlates with disease activity in each disease. It is not a marker specific to SLE. Please discuss from this point of view. We appreciate your critical comments. According to your precise comments, we cited these important references and discussed this point of view in the revised manuscript. ---- Attachments Attachment Submitted filename: Responses to Reviewer Plos one.docx https://doi.org/10.1371/journal.pone.0227069.r002
12 Dec 2019 Decision Letter - Masataka Kuwana, Editor Galectin-9 as a biomarker for disease activity in systemic lupus erythematosus PONE-D-19-27302R1 Dear Dr. Migita, We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements. Within one week, you will receive an e-mail containing information on the amendments required prior to publication. When all required modifications have been addressed, you will receive a formal acceptance letter and your manuscript will proceed to our production department and be scheduled for publication. Shortly after the formal acceptance letter is sent, an invoice for payment will follow. To ensure an efficient production and billing process, please log into Editorial Manager at https://www.editorialmanager.com/pone/, click the "Update My Information" link at the top of the page, and update your user information. 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 enable them to help maximize its impact. If they will be preparing press materials for this manuscript, you must inform our press team as soon as possible and 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. With kind regards, Masataka Kuwana, MD, PhD 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 #2: (No Response) ******** 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 #2: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #2: 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 #2: 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 #2: 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 #2: (No Response) ****** 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 #2: No https://doi.org/10.1371/journal.pone.0227069.r003
Formally Accepted
6 Jan 2020 Acceptance Letter - Masataka Kuwana, Editor PONE-D-19-27302R1 Galectin-9 as a biomarker for disease activity in systemic lupus erythematosus Dear Dr. Migita: I am 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 notify them about your upcoming paper at this point, to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, 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. For any other questions or concerns, please email plosone@plos.org. Thank you for submitting your work to PLOS ONE. With kind regards, PLOS ONE Editorial Office Staff on behalf of Prof. Masataka Kuwana Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0227069.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 .