Peer Review History

Original SubmissionAugust 5, 2023
Decision Letter - Matteo Bauckneht, Editor

PONE-D-23-24585Evaluation of pharmacokinetics, safety, and efficacy of At-211 meta-astatobenzylguanidine (MABG) in patients with pheochromocytoma or paraganglioma (PPGL): A study protocolPLOS ONE

Dear Dr. Kobayakawa,

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 Nov 25 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,

Matteo Bauckneht

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

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 stated that you will provide repository information for your data at acceptance. Should your manuscript be accepted for publication, we will hold it until you provide the relevant accession numbers or DOIs necessary to access your data. If you wish to make changes to your Data Availability statement, please describe these changes in your cover letter and we will update your Data Availability statement to reflect the information you provide.

3. 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.

4. 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.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Does the manuscript provide a valid rationale for the proposed study, with clearly identified and justified research questions?

The research question outlined is expected to address a valid academic problem or topic and contribute to the base of knowledge in the field.

Reviewer #1: Yes

********** 

2. Is the protocol technically sound and planned in a manner that will lead to a meaningful outcome and allow testing the stated hypotheses?

The manuscript should describe the methods in sufficient detail to prevent undisclosed flexibility in the experimental procedure or analysis pipeline, including sufficient outcome-neutral conditions (e.g. necessary controls, absence of floor or ceiling effects) to test the proposed hypotheses and a statistical power analysis where applicable. As there may be aspects of the methodology and analysis which can only be refined once the work is undertaken, authors should outline potential assumptions and explicitly describe what aspects of the proposed analyses, if any, are exploratory.

Reviewer #1: Yes

********** 

3. Is the methodology feasible and described in sufficient detail to allow the work to be replicable?

Descriptions of methods and materials in the protocol should be reported in sufficient detail for another researcher to reproduce all experiments and analyses. The protocol should describe the appropriate controls, sample size calculations, and replication needed to ensure that the data are robust and reproducible.

Reviewer #1: Yes

********** 

4. Have the authors described where all data underlying the findings will be made available when the study is complete?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception, at the time of publication. 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 and, if applicable, provide comments about issues authors must address before this protocol can be accepted for publication. You may also include additional comments for the author, including concerns about research or publication ethics.

You may also provide optional suggestions and comments to authors that they might find helpful in planning their study.

(Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: The work by Kobayakawa and colleagues illustrates the protocol of an ongoing study (started in September 2022) testing the efficacy of [211At]MABG as a therapeutic approach in adult patients (>20 years old) affected by pheochromocytoma/paraganglioma not amenable to surgery.

The study is interesting in light of the paucity of the currently available treatments in this setting and the potential to be transferred to other guanidine-avid diseases, such as neuroblastoma.

I suggest the following amendments to the text:

Please be consistent with the verbal tenses: over the manuscript, you switch from the present to the future tense repeatedly. Since the study is ongoing, you might want to stick to the present tense. Overall, the text reads fine; a review by a native English speaker might be, however, considered.

Please utilise the proper radiopharmaceuticals nomenclature, as indicated in the official EANM statement: https://www.eanm.org/content-eanm/uploads/2019/12/EANM_GUIDANCE-_TRACER_NOMENCLATURE-1.pdf

Please adjust the definition of “multiple active cancers”, which could be confusing in its current form; I suggest “presence of malignancies other than the one object of the study or history of other malignancies with a disease-free interval <5 years”

Please explain which drugs fall in the category described in the second point of the exclusion criteria; the recommended withdrawal period for these medications and the one mentioned in point 3 should also be added.

Please expand on the “uncontrolled thyroid dysfunction” that could lead to an exclusion from the study.

Discussion: consider changing the word “penetrability” to “shorter range in water”; the sentence “have a shorter half-life of At-211” referred to alpha-radiation is confusing and must be corrected or removed.

********** 

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: 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.0303623.r001
Revision 1
Author Response

Responses to Reviewer’s

Comment 1. Does the manuscript provide a valid rationale for the proposed study, with clearly identified and justified research questions?

The research question outlined is expected to address a valid academic problem or topic and contribute to the base of knowledge in the field.

Response to Comment 1: Thank you for your valuable comment. This study is a first-in-human clinical trial of an anticancer drug. The rationale of the study is only from non-clinical pharmacological and biodistribution studies, which indicate the efficacy profiles for pheochromocytoma cells. They are described in the Introduction section (references: 12–17). We have added that the required toxicity studies were conducted prior to the first dose in humans, in accordance with ICH-M3 and -S9 guidelines (https://www.pmda.go.jp/files/000156321.pdf ). Based on these guidelines, we set the start dose for first administration in human, which is described in the Discussion section.

Comment 2. Is the protocol technically sound and planned in a manner that will lead to a meaningful outcome and allow testing the stated hypotheses?

The manuscript should describe the methods in sufficient detail to prevent undisclosed flexibility in the experimental procedure or analysis pipeline, including sufficient outcome-neutral conditions (e.g. necessary controls, absence of floor or ceiling effects) to test the proposed hypotheses and a statistical power analysis where applicable. As there may be aspects of the methodology and analysis which can only be refined once the work is undertaken, authors should outline potential assumptions and explicitly describe what aspects of the proposed analyses, if any, are exploratory.

Response to Comment 2: This study was planned as a phase I study of an anticancer drug. The main purpose of a phase I cancer clinical trial is to determine the maximum tolerated dose and recommended dose by elucidating the dose-limiting toxicity in each dose escalation cohort. That is why, in the present study, there were no statistical hypotheses, and no statistical tests were planned. Regarding exploratory efficacy endpoints, such as objective response and progression-free survival, and pharmacological parameters, point estimations of these parameters are calculated.

Comment 3. Is the methodology feasible and described in sufficient detail to allow the work to be replicable?

Descriptions of methods and materials in the protocol should be reported in sufficient detail for another researcher to reproduce all experiments and analyses. The protocol should describe the appropriate controls, sample size calculations, and replication needed to ensure that the data are robust and reproducible.

Response to Comment 3: Sample size is not calculated statistically, but is traditionally set according to the modified Fibonacci dose escalation (3 + 3 design). In addition, there are usually no controls in phase I cancer clinical trials. Therefore, we have now added an explanation about the modified Fibonacci method in the section describing sample size and statistical analysis. We believe this manuscript ensures reproducibility of study planning and conducting.

Comment 4. Have the authors described where all data underlying the findings will be made available when the study is complete?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception, at the time of publication. 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.

Response to Comment 4: We have added the following sentence to the data availability subsection in the METADATA: “No datasets were generated or analyzed during the current study. All relevant data from this study will be made available upon study completion.”

Comment 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.

Response to Comment 5: We have checked this revised manuscript prior to resubmission, and had it proofread by native English-speaking scientific editors.

Comment 6.

Comment 6-1: Please be consistent with the verbal tenses: over the manuscript, you switch from the present to the future tense repeatedly. Since the study is ongoing, you might want to stick to the present tense. Overall, the text reads fine; a review by a native English speaker might be, however, considered.

Response to Comment 6-1: Thank you for the advice, according to which we have amended the manuscript so that it now uses the correct tenses. In addition, as mentioned in our previous response, the manuscript has also been proofread by native-English-speaking scientific editors.

Comment 6-2: Please utilize the proper radiopharmaceuticals nomenclature, as indicated in the official EANM statement: https://www.eanm.org/content-eanm/uploads/2019/12/EANM_GUIDANCE-_TRACER_NOMENCLATURE-1.pdf

Response to Comment 6-2: We have changed the radiopharmaceuticals’ nomenclature, in accordance with the EANM statement, from At-211 MABG, I-123 MIBG, and I-131 MIBG to [211 At] MABG, [123I] MIBG, [131I] MIBG, respectively.

Comment 6-3: Please adjust the definition of “multiple active cancers”, which could be confusing in its current form; I suggest “presence of malignancies other than the one object of the study or history of other malignancies with a disease-free interval <5 years”

Response to Comment 6-3: Thank you for your suggestion. We have now changed the sentence as follows: ““Multiple active cancers” refers to presence of malignancies other than the one included in the study or history of other malignancies with a disease-free interval <5 years.”

Comment 6-4: Please explain which drugs fall in the category described in the second point of the exclusion criteria; the recommended withdrawal period for these medications and the one mentioned in point 3 should also be added.

Response to Comment 6-4: We have now provided this information in the manuscript as follows: “Patients who cannot stop the dosage of a drug that suppresses the accumulation of MABG according to the EANM procedure guidelines for 131I-meta-iodobenzylguanidine (131I-mIBG) therapy during the study period.” In addition, we have clarified the withdrawal periods of 2 and 3 point as "during the study period."

Comment 6-5: Please expand on the “uncontrolled thyroid dysfunction” that could lead to an exclusion from the study.

Response to Comment 6-5: We have now changed “thyroid dysfunction” to “thyroid dysfunction including hypothyroidism and hyperthyroidism”.

Comment 6-6: Discussion: consider changing the word “penetrability” to “shorter range in water”; the sentence “have a shorter half-life of At-211” referred to alpha-radiation is confusing and must be corrected or removed.

Response to Comment 6-6: Accordingly, we have changed the word “penetrability” to “shorter range in water”, and revised the sentence as follows:

“[211At] At is an α-emitting nuclide with short half-life, and because α-rays have shorter range in water than β-rays, there is no need to isolate patients treated with this drug for long periods in an isotope treatment room.”

Attachments
Attachment
Submitted filename: renamed_c01a4.docx
https://doi.org/10.1371/journal.pone.0303623.r002
Decision Letter - Margo Dona, Editor

PONE-D-23-24585R1Evaluation of pharmacokinetics, safety, and efficacy of [211At] meta-astatobenzylguanidine ([211At] MABG) in patients with pheochromocytoma or paraganglioma (PPGL): A study protocolPLOS ONE

Dear Dr. Kobayakawa,

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 Mar 17 2024 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,

Margo Dona

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.

Additional Editor Comments: Please address the following points of reviewer 2:Include a comprehensive statistical analysis plan. Include details for determining the MTD and RD. Also, state the statistical methods that will be used to analyze the secondary endpoints. Identify the software that will be used to capture the data as well as the software that will be used for the statistical analysis.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Does the manuscript provide a valid rationale for the proposed study, with clearly identified and justified research questions?

The research question outlined is expected to address a valid academic problem or topic and contribute to the base of knowledge in the field.

Reviewer #1: Yes

Reviewer #2: Yes

**********

2. Is the protocol technically sound and planned in a manner that will lead to a meaningful outcome and allow testing the stated hypotheses?

The manuscript should describe the methods in sufficient detail to prevent undisclosed flexibility in the experimental procedure or analysis pipeline, including sufficient outcome-neutral conditions (e.g. necessary controls, absence of floor or ceiling effects) to test the proposed hypotheses and a statistical power analysis where applicable. As there may be aspects of the methodology and analysis which can only be refined once the work is undertaken, authors should outline potential assumptions and explicitly describe what aspects of the proposed analyses, if any, are exploratory.

Reviewer #1: Yes

Reviewer #2: Yes

**********

3. Is the methodology feasible and described in sufficient detail to allow the work to be replicable?

Descriptions of methods and materials in the protocol should be reported in sufficient detail for another researcher to reproduce all experiments and analyses. The protocol should describe the appropriate controls, sample size calculations, and replication needed to ensure that the data are robust and reproducible.

Reviewer #1: Yes

Reviewer #2: No

**********

4. Have the authors described where all data underlying the findings will be made available when the study is complete?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception, at the time of publication. 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

**********

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

Reviewer #2: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above and, if applicable, provide comments about issues authors must address before this protocol can be accepted for publication. You may also include additional comments for the author, including concerns about research or publication ethics.

You may also provide optional suggestions and comments to authors that they might find helpful in planning their study.

(Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Thank you for addressing my comments. I have no further concerns against publication.

Reviewer #2: In this protocol, a phase I clinical trial is currently underway with 3 + 3 dose escalation design to evaluate the pharmacokinetics, safety, and efficacy of [211At] MABG at 3 dose levels in patients with unresectable or metastatic PPGI. The primary endpoint is dose-limiting toxicity to determine the maximum tolerated dose and recommended doses. The secondary endpoints include radiopharmacokinetics, urinary radioactive excretion rate, urinary catecholamine response rate, objective response rate, progression free survival, [123I] MIBG scintigraphy on reducing tumor accumulation, and quality of life.

Major revisions:

Include a comprehensive statistical analysis plan. Include details for determining the MTD and RD. Also, state the statistical methods that will be used to analyze the secondary endpoints.

Minor revisions:

1-Identify the software that will be used to capture the data as well as the software that will be used for the statistical analysis.

2-To assist in the review process, add line numbering to the document.

**********

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: 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.0303623.r003
Revision 2
Author Response

Responses to Reviewer’s

Comment of Reviewer 2.

Major revisions: Include a comprehensive statistical analysis plan. Include details for determining the MTD and RD. Also, state the statistical methods that will be used to analyze the secondary endpoints.

Minor revisions. 1-Identify the software that will be used to capture the data as well as the software that will be used for the statistical analysis. 2-To assist in the review process, add line numbering to the document.

Response to all Comments of Reviewer 2: Thanks for your valuable comments. We have added following sentences to the “Statistical analysis” section (pages 20 to 21):

“MTD is determined as the highest dose level at which none of the three cases developed DLT or only one of the six cases developed DLT. RD is determined as the same dose of MTD. All secondary endpoints are described as descriptive statistics, no formal statistical hypothesis test is performed.　Statistical analyses are performed with SAS software, version 9.4 (SAS Institute).”

“PK/PD analyses are performed with Phoenix WinNonlin software (version 8.1, Certara, Inc.).”

Attachments
Attachment
Submitted filename: renamed_657e6.docx
https://doi.org/10.1371/journal.pone.0303623.r004
Decision Letter - Margo Dona, Editor

Evaluation of pharmacokinetics, safety, and efficacy of [211At] meta-astatobenzylguanidine ([211At] MABG) in patients with pheochromocytoma or paraganglioma (PPGL): A study protocol

PONE-D-23-24585R2

Dear Dr. Kobayakawa,

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 will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. If you have any questions relating to publication charges, 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,

Margo Dona

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Does the manuscript provide a valid rationale for the proposed study, with clearly identified and justified research questions?

The research question outlined is expected to address a valid academic problem or topic and contribute to the base of knowledge in the field.

Reviewer #2: Yes

**********

2. Is the protocol technically sound and planned in a manner that will lead to a meaningful outcome and allow testing the stated hypotheses?

The manuscript should describe the methods in sufficient detail to prevent undisclosed flexibility in the experimental procedure or analysis pipeline, including sufficient outcome-neutral conditions (e.g. necessary controls, absence of floor or ceiling effects) to test the proposed hypotheses and a statistical power analysis where applicable. As there may be aspects of the methodology and analysis which can only be refined once the work is undertaken, authors should outline potential assumptions and explicitly describe what aspects of the proposed analyses, if any, are exploratory.

Reviewer #2: Yes

**********

3. Is the methodology feasible and described in sufficient detail to allow the work to be replicable?

Descriptions of methods and materials in the protocol should be reported in sufficient detail for another researcher to reproduce all experiments and analyses. The protocol should describe the appropriate controls, sample size calculations, and replication needed to ensure that the data are robust and reproducible.

Reviewer #2: Yes

**********

4. Have the authors described where all data underlying the findings will be made available when the study is complete?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception, at the time of publication. 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: No

**********

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 and, if applicable, provide comments about issues authors must address before this protocol can be accepted for publication. You may also include additional comments for the author, including concerns about research or publication ethics.

You may also provide optional suggestions and comments to authors that they might find helpful in planning their study.

(Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #2: All comments have been adequately addressed.

**********

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.0303623.r005
Formally Accepted
Acceptance Letter - Margo Dona, Editor

PONE-D-23-24585R2

PLOS ONE

Dear Dr. Kobayakawa,

I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now being handed over to our production team.

At this stage, our production department will prepare your paper for publication. This includes ensuring the following:

* All references, tables, and figures are properly cited

* All relevant supporting information is included in the manuscript submission,

* There are no issues that prevent the paper from being properly typeset

If revisions are needed, the production department will contact you directly to resolve them. If no revisions are needed, you will receive an email when the publication date has been set. At this time, we do not offer pre-publication proofs to authors during production of the accepted work. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few weeks to review your paper and let you know the next and final steps.

Lastly, 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 customercare@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. Margo Dona

Academic Editor

PLOS ONE

https://doi.org/10.1371/journal.pone.0303623.r006

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 .