Non-invasive intradermal imaging of cystine crystals in cystinosis

Marya Bengali; Spencer Goodman; Xiaoying Sun; Magdalene A. Dohil; Ranjan Dohil; Robert Newbury; Tatiana Lobry; Laura Hernandez; Corinne Antignac; Sonia Jain; Stephanie Cherqui

doi:10.1371/journal.pone.0247846

Peer Review History

Original SubmissionOctober 5, 2020
14 Jan 2021 Decision Letter - Thomas Abraham, Editor Transfer Alert This paper was transferred from another journal. As a result, its full editorial history (including decision letters, peer reviews and author responses) may not be present. PONE-D-20-31230 Non-invasive intradermal imaging of cystine crystals in cystinosis PLOS ONE Dear Dr. Cherqui, 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 Feb 28 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. 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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: This study highlights promising new technology to non-invasively follow disease status in cystinosis. There are no previous studies done with this technology for cystinosis patient. Manuscript is technically sound and data support the conclusion but below are some comments: 1. Include final sample sizes used in statistics section within methods. For example it was not clear what sample size was used for ELISA? Mention if replicates were performed? 2. Point out in H&E staining image perivascular chronic inflammatory cells in figure 1A as mentioned in page 10. 3. Figure 2 C states that “Preliminary slices were excluded due to signal background these skin layers in healthy controls.” This statement is not clear, clarify this statement further, What is the tissue depth of these excluded slices? Were they present in the starred region? What is justification to remove them? Will this effect results if we include them, if yes is there background in patients’ images as well at this location? By excluding this are we still comparing pateints vs controls in similar experimental setup/ conditions? 4. Indicate sample size used for figure 3D 5. Typographical error in page 14 second paragraph line 3. Correct hrough to “Through” 6. Conclusion is based on results observed but can this methodology be applied to people with with dark skin people? Are there any other challenges with this methodology? ****** 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 [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: Plos One.pdf https://doi.org/10.1371/journal.pone.0247846.r001
Revision 1
25 Jan 2021 Author Response Response to Academic Editor: 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. Response: we formatted the revised manuscript to meet PLOS ONE's style requirements. 2. Please provide additional details regarding participant consent. In the ethics statement in the Methods and online submission information, please ensure that you have specified what type you obtained (for instance, written or verbal, and if verbal, how it was documented and witnessed). If your study included minors, state whether you obtained consent from parents or guardians. If the need for consent was waived by the ethics committee, please include this information. Response: This information has been added in the Methods section page 6. 3.We note that you have a patent relating to material pertinent to this article. Please provide an amended statement of Competing Interests to declare this patent (with details including name and number), along with any other relevant declarations relating to employment, consultancy, patents, products in development or modified products etc. Please confirm that this does not alter your adherence to all PLOS ONE policies on sharing data and materials, as detailed online in our guide for authors http://journals.plos.org/plosone/s/competing-interests by including the following statement: "This does not alter our adherence to PLOS ONE policies on sharing data and materials.” If there are restrictions on sharing of data and/or materials, please state these. Please note that we cannot proceed with consideration of your article until this information has been declared. 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 Response: I updated my disclosure with all my patent information. However, these patents are not related to this article. The patent entitled “Methods of treating mitochondrial disorders” and “Methods of treating lysosomal disorders” are related to treating mitochondrial and lysosomal diseases, respectively, with gene-corrected hematopoietic stem cells. The present study reports a new technology to non-invasively follow-up the disease status of patients affected with cystinosis. There is no patent for this present study so no restriction for sharing the data or materials. 4. Your ethics statement should only appear in the Methods section of your manuscript. If your ethics statement is written in any section besides the Methods, please delete it from any other section. Response: The ethics statement only appears in the Methods section. 5. Please upload a copy of Supporting Information eVideos 1-2., eVideos 3-4., eVideos 5-6. and eVideos 7-8.which you refer to in your text on page 26. Response: While we thought these files were uploaded, we apologize if they were not. The Supporting Information eVideos, now renamed as S# Videos, have been uploaded. 6. Please upload a copy of eFigure 1, eFigure 2A, eFigure 3 and eFigure 4 to which you refer in your text on page 7, 9, 11 and 12. If the figure is no longer to be included as part of the submission please remove all reference to it within the text. Response: While we thought these files were uploaded, we apologize if they were not. The Supporting Information file containing the eFigures, now renamed as S# Figures, has been uploaded. Response to Reviewers This study highlights promising new technology to non-invasively follow disease status in cystinosis. There are no previous studies done with this technology for cystinosis patient. Manuscript is technically sound and data support the conclusion but below are some comments: Response: We acknowledge the reviewer for his comments. 1. Include final sample sizes used in statistics section within methods. For example it was not clear what sample size was used for ELISA? Mention if replicates were performed? Response: We added this information in the ELISA section in the Methods page 7. Also, this information also appears in the S2 Figure legend. Finally, in the Supporting Information, which seems to be missing in the final submission, there is the S1 Table that details the numbers of patients and controls enrolled and analyzed by confocal microscope and the number of medical records obtained. 2. Point out in H&E staining image perivascular chronic inflammatory cells in figure 1A as mentioned in page 10. Response: Arrowhead has been added in Figure 1A to point out the perivascular chronic inflammatory cells. 3. Figure 2 C states that “Preliminary slices were excluded due to signal background these skin layers in healthy controls.” _This statement is not clear, clarify this statement further, What is the tissue depth of these excluded slices? Were they present in the starred region? What is justification to remove them? Will this effect results if we include them, if yes is there background in patients_’ _images as well at this location? By excluding this are we still comparing pateints vs controls in similar experimental setup/ conditions? Response: We thank the reviewer to have pointed out this sentence, which was actually a mistake as we did not exclude any slice for the 2D quantification. Indeed, as stipulated in the Result section “Crystal quantification by automated image analysis in 2D”, out of the 78 slices that were captured per patient, significantly higher total crystal area was observed in patients compared to controls within the papillary dermis region, corresponding to slices 15-50 (Fig 2C, S3 Fig., S4 Video). The differences were not significant in the epidermis and hypodermis because of high background and low clarity, respectively (S2 Table). Therefore, for the 3D quantification we used only the papillary dermis region which present the least artifacts and background. We corrected the Figure 2 legend accordingly. 4. Indicate sample size used for figure 3D Response: Each patient is represented by a dot; we added this information in the legend of figure 3D as well as sample size for each medical outcome. This information is also available in the complete dataset supplementary Table “2019 Biostasts Complete Dataset”. 5. Typographical error in page 14 second paragraph line 3. Correct hrough to “Through” _ Response: This typo has been corrected. 6. Conclusion is based on results observed but can this methodology be applied to people with with dark skin people? Are there any other challenges with this methodology? Response: As stipulated in the manuscript, people with dark skin will present with higher background level, especially in the epidermis and hypodermis area. This is indeed a limitation of this technology. However, for the 3D quantification of the cystine crystal, we use only the papillary dermis region, which mitigate this issue. In addition, this technology is specific for imaging and quantifying cystine crystal in the skin, which can be found only in cystinosis patients who have very pale skin due to deficiency in melanin production [1]. Therefore, this issue is very limited for this particular population. We made this point clearer in the results section page 11. 1. Chiaverini C, Sillard L, Flori E, Ito S, Briganti S, Wakamatsu K, et al. Cystinosin is a melanosomal protein that regulates melanin synthesis. FASEB journal : official publication of the Federation of American Societies for Experimental Biology. 2012;26(9):3779-89. doi: 10.1096/fj.11-201376. PubMed PMID: 22649030. https://doi.org/10.1371/journal.pone.0247846.r002
15 Feb 2021 Decision Letter - Thomas Abraham, Editor Non-invasive intradermal imaging of cystine crystals in cystinosis PONE-D-20-31230R1 Dear Dr. Cherqui, 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, Thomas Abraham, PhD Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0247846.r003
Formally Accepted
23 Feb 2021 Acceptance Letter - Thomas Abraham, Editor PONE-D-20-31230R1 Non-invasive intradermal imaging of cystine crystals in cystinosis Dear Dr. Cherqui: 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. Thomas Abraham Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0247846.r004

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