A potential diagnostic serum immunological marker panel to differentiate between primary and secondary knee osteoarthritis

Sakuni Rankothgedera; Inoshi Atukorala; Chandrika Fernando; Duminda Munidasa; Lalith Wijayaratne; Preethi Udagama

doi:10.1371/journal.pone.0257507

Peer Review History

Original SubmissionApril 18, 2021
6 Jun 2021 Decision Letter - Oreste Gualillo, Editor PONE-D-21-12809 A potential diagnostic serum immunological marker panel to differentiate between primary and secondary knee osteoarthritis PLOS ONE Dear Dr. Udagama, 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 Jul 21 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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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. We note that you have indicated that data from this study are available upon request. PLOS only allows data to be available upon request if there are legal or ethical restrictions on sharing data publicly. For information on unacceptable data access restrictions, please see http://journals.plos.org/plosone/s/data-availability#loc-unacceptable-data-access-restrictions. In your revised cover letter, please address the following prompts: 2a) If there are ethical or legal restrictions on sharing a de-identified data set, please explain them in detail (e.g., data contain potentially identifying or sensitive patient information) and who has imposed them (e.g., an ethics committee). Please also provide contact information for a data access committee, ethics committee, or other institutional body to which data requests may be sent. 2b) If there are no restrictions, please upload the minimal anonymized data set necessary to replicate your study findings as either Supporting Information files or to a stable, public repository and provide us with the relevant URLs, DOIs, or accession numbers. Please see http://www.bmj.com/content/340/bmj.c181.long for guidelines on how to de-identify and prepare clinical data for publication. For a list of acceptable repositories, please see http://journals.plos.org/plosone/s/data-availability#loc-recommended-repositories. We will update your Data Availability statement on your behalf to reflect the information you provide. Additional Editor Comments : Dear Authors, after careful revision of the article and considering the reviewers comments, my decision as academic editor is "Major Revision". Both reviewers have recognized clear interest in the article and I think too that your article is in line with PLOSONE criteria. However, in the current form there are certain aspects that, for the time being, preclude the acceptance of the article. I suggest to follow point by point all the suggestions proposed by the reviewers. You have granted standard time for the review, but if you need extra time, please do not hesitate in asking for it. Thank you for your contribution to PLOSONE. Dr. Oreste Gualillo, PharmD, PhD PLOSONE Academic Editor. [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: Yes Reviewer #2: Partly ******** 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: No ****** 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: In this study, Rankothgedera S. et al aimed to identify potential serum immunologic diagnostic markers of primary and secondary knee osteoarthritis (pKOA and sKOA, respectively). Their results suggest that there might be distinct panels of markers for pKOA and sKOA that, if confirmed, may be useful for the development of therapeutic strategies for these two types of KOA. The study is well designed, the methodology is appropriate and the conclusions are supported by the results obtained. However, there are some major issues that should be resolved prior to the acceptance of the manuscript: 1) The authors should define the abbreviations upon their first appearance in the text, and always use the same abbreviation (e.g. when talking about primary and secondary knee osteoarthritis, sometimes they refer to it as pKOA or sKOA, sometimes as primary/secondary OA). Please unify. 2) Additionally, after an abbreviation is defined for the first time, they should use the abbreviation thenceforth (e.g. RA, SLE). 3) In the Statistical Analyses sub-section the authors should detail which were the confounding factors included in the analysis. 4) In the tables the authors should define the abbreviations used in the legends, below them. 5) Figure 1 A and B: The type of graph chosen by the authors is not correct for the data/results they want to show. They should use a box plot or scatter plot, for example. But they should not add connecting lines between the values obtained in each study group for each marker. 6) In the Statistical Analyses sub-section the authors mention that the level of statistical significance was set at p<0.05, meaning that a p-value of 0.05 is not considered as statistically significant. However, some of the results reported in this manuscript as statistically significant exhibit a p-value = 0.05. Please check and correct. 7) The number of individuals with sKOA is relatively low. Accordingly, this should also be included as a potential limitation of the study, since the low sample size of this group may affect the results obtained (and the conclusions drawn from them). In addition, there are some minor issues that should also be addressed: 1) Line 164: Some punctuation marks are missing in the caption of Figure 1. Please check. 2) Line 176: The correct phrase is ‘on par’, not ‘in par’. 3) Line 186: Please, cite Figure 2 at the end of the paragraph. 4) Line 242: The authors should replace ‘Females’ by ‘Female sex’. 5) Line 247: The term ‘that’ is repeated. Reviewer #2: In the manuscript titled “A potential diagnostic serum immunological marker panel to differentiate between primary and secondary knee osteoarthritis” the authors identified a serum biomarker panels able to differentiate primary from secondary knee osteoarthritis (KOA). The paper includes an adequate statistical analysis and provides experimental findings that allow to distinguish primary OA from secondary disease. However, there are some aspects, that should be considered to improve the overall completeness and quality of the paper. 1. It is not clear to this referee how the study was designed ; specifically, it is no specified timing of blood withdraw from KOA patients. Also, disease stage was not indicated. Please, consider these aspects also for SLE subjects. This are very important issues in order to determine the “predictive” role of the panel proposed. 2. In this context, the authors should provide a schematic diagram representing the study design. The authors investigated the of Th1/Th2 ratio on KOA and analyzed TNF�:IL-10 ratio as readout of Th1/Th2 balance. However, other cytokines contribute to this balance (e.g, IFN-a, IL-2, IL-12 Vs IL-4, IL-5); some of these additional cytokines should be measured to better define Th1/Th2 balance. Throughout the manuscript the data are not well presented. As relevant examples, figure 1 (A & B) and figure 2 are not clear. In this context, box plot with dots should be used to improve the message from these findings. ****** 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.] 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. https://doi.org/10.1371/journal.pone.0257507.r001
Revision 1
23 Aug 2021 Author Response Responses to reviewers’ comments: PONE-D-21-12809: A potential diagnostic serum immunological marker panel to differentiate between Primary and Secondary Knee Osteoarthritis As coauthors of this manuscript, we deeply appreciate the constructive comments and suggestions made by the reviewers; we have attempted to address all queries raised by the two reviewers. We hope our manuscript now meets the caliber of the journal and will be suitable to be published in PLOS ONE. Reviewer #1: In this study, Rankothgedera S. et al aimed to identify potential serum immunologic diagnostic markers of primary and secondary knee osteoarthritis (pKOA and sKOA, respectively). Their results suggest that there might be distinct panels of markers for pKOA and sKOA that, if confirmed, may be useful for the development of therapeutic strategies for these two types of KOA. The study is well designed, the methodology is appropriate and the conclusions are supported by the results obtained. However, there are some major issues that should be resolved prior to the acceptance of the manuscript: 1) The authors should define the abbreviations upon their first appearance in the text, and always use the same abbreviation (e.g. when talking about primary and secondary knee osteoarthritis, sometimes they refer to it as pKOA or sKOA, sometimes as primary/secondary OA). Please unify. The suggested changes were made in the revised manuscript. 2) Additionally, after an abbreviation is defined for the first time, they should use the abbreviation thenceforth (e.g. RA, SLE). The original manuscript was revised based on the reviewer comment. 3) In the Statistical Analyses sub-section the authors should detail which were the confounding factors included in the analysis. Confounding factors used in the analysis were included in the Statistical Analyses sub-section of the revised manuscript (lines 150-151). 4) In the tables the authors should define the abbreviations used in the legends, below them. This shortcoming in Tables 1 to 4 was corrected in the revised manuscript. 5) Figure 1 A and B: The type of graph chosen by the authors is not correct for the data/results they want to show. They should use a box plot or scatter plot, for example. But they should not add connecting lines between the values obtained in each study group for each marker. Taking into consideration the reviewer’s comment, Figures 1 and 2 (Figs 2 and 3 in the revised manuscript) are presented as box plots (with dots), in the revised manuscript. 6) In the Statistical Analyses sub-section the authors mention that the level of statistical significance was set at p<0.05, meaning that a p-value of 0.05 is not considered as statistically significant. However, some of the results reported in this manuscript as statistically significant exhibit a p-value = 0.05. Please check and correct. In the Statistical Analyses sub – section, the last sentence in the revised manuscript reads as “The level of statistical significance was set at p≤0.05” (line 154). 7) The number of individuals with sKOA is relatively low. Accordingly, this should also be included as a potential limitation of the study, since the low sample size of this group may affect the results obtained (and the conclusions drawn from them). The sentence, “Nonetheless, the low sample size of the sKOA group may present as a potential limitation of this study, which may perhaps have ramifications (lines 289-291)” was inserted. In addition, there are some minor issues that should also be addressed: 1) Line 164: Some punctuation marks are missing in the caption of Figure 1. Please check. 2) Line 176: The correct phrase is ‘on par’, not ‘in par’. 3) Line 186: Please, cite Figure 2 at the end of the paragraph. 4) Line 242: The authors should replace ‘Females’ by ‘Female sex’. 5) Line 247: The term ‘that’ is repeated. The aforementioned minor corrections were rectified in the revised manuscript. Reviewer #2: In the manuscript titled “A potential diagnostic serum immunological marker panel to differentiate between primary and secondary knee osteoarthritis” the authors identified a serum biomarker panels able to differentiate primary from secondary knee osteoarthritis (KOA). The paper includes an adequate statistical analysis and provides experimental findings that allow to distinguish primary OA from secondary disease. However, there are some aspects that should be considered to improve the overall completeness and quality of the paper. 1. It is not clear to this referee how the study was designed; specifically, it is no specified timing of blood withdrawal from KOA patients. Also, disease stage was not indicated. Please, consider these aspects also for SLE subjects. These are very important issues in order to determine the “predictive” role of the panel proposed. In this context, the authors should provide a schematic diagram representing the study design. The following revisions were incorporated to the revised manuscript: Under Materials and Methods, sub section ‘Study Participants’ • The case control study design (scheme) was incorporated as Figure 1 (lines 102-103), • Both pKOA (line 112) and sKOA (line 116) patients recruited for the study were at Stage 3 of the disorders. • All SLE patients recruited as disease controls for the study, specifically manifested lupus arthritis (line 109-110). . 2. The authors investigated the Th1/Th2 ratio on KOA and analyzed TNF�:IL-10 ratio as readout of Th1/Th2 balance. However, other cytokines contribute to this balance (e.g, IFN-a, IL-2, IL-12 Vs IL-4, IL-5); some of these additional cytokines should be measured to better define Th1/Th2 balance. While completely agreeing with the reviewer on this viewpoint, lack of funds restricted us to assay selected serum cytokines. Hence, the sentences “However, the assay of additional serum cytokines such as IFN-γ, IL-2, IL-12 Vs IL-4, IL-5 would have explicitly defined the Th1:Th2 balance among the study groups. This was not pragmatic due to restricted funds” were inserted (lines 197-199). 3.Throughout the manuscript the data are not well presented. As relevant examples, figure 1 (A & B) and figure 2 are not clear. In this context, box plot with dots should be used to improve the message from these findings. Taking into consideration the reviewer’s comment, Figures 1 and 2 (Figs 2 and 3 in the revised manuscript) are presented as box plots (with dots), in the revised manuscript. Attachments Attachment Submitted filename: Response to Reviewers .docx https://doi.org/10.1371/journal.pone.0257507.r002
3 Sep 2021 Decision Letter - Oreste Gualillo, Editor A potential diagnostic serum immunological marker panel to differentiate between Primary and Secondary Knee Osteoarthritis PONE-D-21-12809R1 Dear Dr. Udagama, 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, Oreste Gualillo, PharmD, PhD Academic Editor PLOS ONE Additional Editor Comments (optional): No additional comments. 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: 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 #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: The authors addresse most of the prevoius concerns. I suggest the to remobe the sentence "this was not pragrmatic due the restrict fund". Add the absence of Th1/Th2 characterization as one of the limitations of the study. ****** 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.0257507.r003
Formally Accepted
9 Sep 2021 Acceptance Letter - Oreste Gualillo, Editor PONE-D-21-12809R1 A potential diagnostic serum immunological marker panel to differentiate between Primary and Secondary Knee Osteoarthritis Dear Dr. Udagama: 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 Oreste Gualillo Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0257507.r004

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