Inner hair cell dysfunction in Klhl18 mutant mice leads to low frequency progressive hearing loss

Neil J. Ingham; Navid Banafshe; Clarisse Panganiban; Julia L. Crunden; Jing Chen; Morag A. Lewis; Karen P. Steel

doi:10.1371/journal.pone.0258158

Peer Review History

Original SubmissionJuly 27, 2021
3 Sep 2021 Decision Letter - Olivia Bermingham-McDonogh, Editor PONE-D-21-24337 Inner hair cell dysfunction in Klhl18 mutant mice leads to low frequency progressive hearing loss PLOS ONE Dear Dr. Ingham, 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. The manuscript needs the minor revisions suggested by Reviewer 1. Please submit your revised manuscript by Oct 18 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. Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. 3. Please include a copy of Table S1 which you refer to in your text on page 13, 14, 16, 17, 18, 20, 21, 38, 39, 40 and 41. [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: 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 manuscript from Neil Ingham and co-authors entitled “Inner hair cell dysfunction in Klhl18 mutant mice leads to low frequency progressive hearing loss” is precisely crafted and has an abundance of supporting data that is presented in good-looking figures. It so nice at least once in a while to read a manuscript in near final form with thorough evaluations and properly replicated data. There are only a few minor issues to suggest to the authors. In the abstract and text there are sentences such as “Scanning electron microscopy showed that Klhl18lowf mutant mice had abnormally tapering inner hair cell stereocilia in the apical half of the cochlea while their synapses appeared normal.” The stereocilia taper usually refers to the base of a stereocilium where the stereocilium joins with the apical surface of the hair cell. That’s not the taper the authors are referring to and that becomes obvious when examining the beautiful images provided. The authors should make an effort to better define both in the abstract and in the main text that they are referring to abnormal tapers near the distal end of stereocilia. The first time a variant of Klhl18 is mentioned, indicate if the allele is dominant or recessive. The statement on line 51 “Neither allele produced overt vestibular phenotypes and homozygotes are viable and fertile [1,2].” could be improved by mentioning the vestibular tests. “overt” isn’t very scientific. What vestibular abnormal phenotype would be classified as overt that these mice didn’t exhibit? Reviewer #2: Inner hair cell dysfunction in Klhl18 mutant mice leads to low frequency progressive hearing loss The manuscript deals with characterization of a model for a hearing phenotype that is found in, but less well studied, in the human population. The Klhl18 (Kelch-like family member 18) gene was examined using two different alleles in the mouse. Interestingly, this gene has been implicated in retina function. The morphological changes in the mice were subtle, with normal DPOAE, CM and synaptic contacts. However, the inner hair cells had some morphological changes (tapering in the inner hair cells), backed up by summating potentials, implicating these cells as impaired in the Klhl18 mutants. The analysis is extremely thorough, and provides a mechanism of function for the low frequency progressive hearing loss due to mutations in this gene. It will be interesting to see if human pathogenic variants might turn up if such families will be screened by whole exome sequencing. ****** 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.0258158.r001
Revision 1
17 Sep 2021 Author Response Responses to reviewers’ comments, Ingham et al. 1. When submitting your revision, we need you to address these additional requirements. 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 >> We have edited the manuscript to comply with the journal formatting requirements and tracked all changes. 2. Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. >> We have checked the reference list but have not noticed any publications that we know have been retracted. 3. Please include a copy of Table S1 which you refer to in your text on page 13, 14, 16, 17, 18, 20, 21, 38, 39, 40 and 41. >> We apologize for overlooking the inclusion of Table S1 in the original submission. An Excel file containing Table S1 (named “_TableS1_Ingham et al.xlsx”) is included with this revision. Reviewer #1: The manuscript from Neil Ingham and co-authors entitled “Inner hair cell dysfunction in Klhl18 mutant mice leads to low frequency progressive hearing loss” is precisely crafted and has an abundance of supporting data that is presented in good-looking figures. It so nice at least once in a while to read a manuscript in near final form with thorough evaluations and properly replicated data. >>Thank you for these comments. There are only a few minor issues to suggest to the authors. Klhl18lowf mutant mice had abnormally tapering inner hair cell stereocilia in the apical half of the cochlea while their synapses appeared normal.” The stereocilia taper usually refers to the base of a stereocilium where the stereocilium joins with the apical surface of the hair cell. That’s not the taper the authors are referring to and that becomes obvious when examining the beautiful images provided. The authors should make an effort to better define both in the abstract and in the main text that they are referring to abnormal tapers near the distal end of stereocilia. >> Thank you for pointing out that confusion. We have altered the text at multiple locations to clarify that we are talking about tapering of the tips of stereocilia, not the insertion point into the cuticular plate. The first time a variant of Klhl18 is mentioned, indicate if the allele is dominant or recessive. >> We have added comments in the abstract and introduction that the hearing loss is recessive in inheritance. However, there may be a semi-dominant effect on stereocilia tip tapering. It is the phenotype studied that is recessive or dominant rather than the allele, so we are cautious in the text to describe hearing loss as recessive, not the full phenotype. We found no differences in our measures of auditory function between heterozygotes and wildtypes. The statement on line 51 “Neither allele produced overt vestibular phenotypes and homozygotes are viable and fertile [1,2].” could be improved by mentioning the vestibular tests. “overt” isn’t very scientific. What vestibular abnormal phenotype would be classified as overt that these mice didn’t exhibit? >> We have added “such as circling or head-bobbing” to the text at this point to clarify. Reviewer #2: Inner hair cell dysfunction in Klhl18 mutant mice leads to low frequency progressive hearing loss The manuscript deals with characterization of a model for a hearing phenotype that is found in, but less well studied, in the human population. The Klhl18 (Kelch-like family member 18) gene was examined using two different alleles in the mouse. Interestingly, this gene has been implicated in retina function. The morphological changes in the mice were subtle, with normal DPOAE, CM and synaptic contacts. However, the inner hair cells had some morphological changes (tapering in the inner hair cells), backed up by summating potentials, implicating these cells as impaired in the Klhl18 mutants. The analysis is extremely thorough, and provides a mechanism of function for the low frequency progressive hearing loss due to mutations in this gene. It will be interesting to see if human pathogenic variants might turn up if such families will be screened by whole exome sequencing. >> Thank you for these comments. We are looking out for human mutations as the reviewer suggests. Attachments Attachment Submitted filename: Ingham et al Klhl18 Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0258158.r002
20 Sep 2021 Decision Letter - Olivia Bermingham-McDonogh, Editor Inner hair cell dysfunction in Klhl18 mutant mice leads to low frequency progressive hearing loss PONE-D-21-24337R1 Dear Dr. Ingham, 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, Olivia Bermingham-McDonogh, Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0258158.r003
Formally Accepted
24 Sep 2021 Acceptance Letter - Olivia Bermingham-McDonogh, Editor PONE-D-21-24337R1 Inner hair cell dysfunction in Klhl18 mutant mice leads to low frequency progressive hearing loss Dear Dr. Ingham: 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. Olivia Bermingham-McDonogh Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0258158.r004

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