PONE-D-23-01537Presence of Known Feline ALMS1 and MYBPC3 Variants in a Diverse Cohort of Cats with Hypertrophic Cardiomyopathy in JapanPLOS ONE

Dear Dr. Matsumoto,

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Shinsuke Yuasa

Academic Editor

PLOS ONE

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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

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: No

Reviewer #2: Yes

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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: No

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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

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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: SUMMARY:

The authors investigated the clinical prevalence of HCM gene variants in cats. They reported rare occurrences of MYBPC3 p.A31P and ALMS1 p.G3376R in specific cat breeds which have not been reported yet. Although this report has important implications for the future application of genetic testing in cats, several points must be addressed.

GENERAL COMMENTS:

The contents of the manuscript should be stated in an appropriate order. For example, limitations and conclusions are not listed where they should be. That makes the manuscript confusing. Also, the reviewer highly recommends the authors make subtitles for each subsection in the Discussion, and the first subsection should be the summary of findings in the present study. These modifications would make the manuscript easier to comprehend.

Whereas MYBPC3 p.A31P is an established genetic mutation in HCM, ALMS1 p.G3376R is not established (the association with HCM is just suggested). These two findings should not be listed in the same importance. The authors should state the clinical implication of these findings in each subsection, or in each paper.

The last paragraph of the Introduction contains the results of this investigation. The author should move these descriptions to the discussion section, or conclusions section (create it after the discussion section).

In the last sentence of page 16, the expression “genetic testing of MYBPC3 p.A31P in non-Maine Coon breeds is necessary for HCM diagnosis and to prevent the establishment of HCM-affected colonies”, seems too strong for the suggestion from results with a relatively small number of subjects. The author would be advised to alter the expression in an appropriate manner.

MINOR COMMENTS:

(Discussion, Page 14, Line 13) The author should add “Bell et al., 2012” into the Reference.

(Table 1) As per the manuscript, the MYBPC3 p.A31P in Munchkin is the variant that has never been found. Therefore, an asterisk should be attached next to the number “1”.

(Table 3) There is no row about Minuet. As per the manuscript, one heterozygote ALMS1 variant was found in the 2 Minuet cats. If the author considers the ALMS1 variant in Minuet is worth reporting, it should be listed also in the Table.

Figure 1 seems not to have enough information. It should be transformed into another table, which enables the authors to fill out the absolute numbers of each zygosity in each group. They can also attach percentages next to each absolute number.

Reviewer #2: The authors of the article present the results of genetic study of five variants previously associated with HCM in 57 HCM-affected, 19 HCM-unaffected, and 227 non-examined cats from the Japanese population to investigate the occurrence among breeds. They conclude that two of these variants could not be limited to specific breeds, as previously it has been reported.

- Abstract

You should check the variant name p.R830W.

- Introduction

At the end of this section you describe your results and conclusion. You shouldn’t include your results in the introduction Section.

- Methods

The authors include 2 Minuet cats without a cardiac phenotype in “Phenotyping and selected animals” section, nevertheless the genetic results are not provided. The authors should include the genetics results of these cats in table 3.

In this section the authors must detail the statistical analysis used.

- Results

I suggest to the authors that they include a table with the main clinical variables of the cats studied such as breed, age, age of phenotype, gender, mm hypertrophy, events or arrhythmia.

The genetic results could be integrated and summarized in a table that includes the frequency of the variants and the comparison between the different groups of animals.

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Reviewer #1: Yes: Keitaro Akita

Reviewer #2: No

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PONE-D-23-01537R1Presence of Known Feline ALMS1 and MYBPC3 Variants in a Diverse Cohort of Cats with Hypertrophic Cardiomyopathy in JapanPLOS ONE

Dear Dr. Matsumoto,

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 Apr 15 2023 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,

Shinsuke Yuasa

Academic Editor

PLOS ONE

Journal Requirements:

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.

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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 #1: 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 #1: Partly

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: 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 #1: 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 #1: 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 #1: The authors have partially addressed the questions and comments raised in the last revision. There are a few points that must be addressed.

The aim of this study was removed from the whole manuscript. It should be retained in the last part of the introduction.

The reviewer still recommends that the authors should state the conclusion at the end of the manuscript. It should the main message to the reader clearer.

All the abbreviations in the supplemental tables should be spelled out in the footnotes.

Although the authors added the clinical information of subjects in the supplemental table 2, they did not describe it in either the results or the discussion in detail. They should describe the clinical characteristics or cardiac events specific to MYBPC3 p.A31p positive cases or ALMS1p.G3376R positive cases.

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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.

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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: Yes: Keitaro Akita

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[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.

Presence of known feline ALMS1 and MYBPC3 variants in a diverse cohort of cats with hypertrophic cardiomyopathy in Japan

PONE-D-23-01537R2

Dear Dr. Matsumoto,

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.

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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,

Shinsuke Yuasa

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 #1: 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 #1: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: 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 #1: 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 #1: 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 #1: The authors have successfully addressed the comments raised in the last revision. The reviewer has no further comments.

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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 #1: Yes: Keitaro Akita

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