PONE-D-21-25364Protocol for chronic hepatitis B virus infection mouse model development by patient-derived orthotopic xenograftsPLOS ONE

Dear Dr. Nagornykh,

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Ezio Laconi, MD, PhD

Academic Editor

PLOS ONE

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 “This work is supported by the Ministry of Science and Higher Education of the Russian Federation within the framework of a grant in the form of a subsidy for the creation and development of the «World-class Genomic Research Center for Ensuring Biological Safety and Technological Independence under the Federal Scientific and Technical Program for the Development of Genetic Technologies», agreement No. 075-15-2019-1666.

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4. Please amend the manuscript submission data (via Edit Submission) to include author Vasiliy G. Akimkin

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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: No

Reviewer #2: Partly

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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: Partly

Reviewer #2: No

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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: No

Reviewer #2: No

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

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Reviewer #1: No

Reviewer #2: No

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Reviewer #1: No

Reviewer #2: No

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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.

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

Reviewer #1: Aleksey et al tried to compare and establish an optimized protocol to generate HBV-HCC PDX mouse model. However, the current version needs to be improved.

Major comment,

1. No any realy result is shown. Please provide your own data.

2. Authors should illustrate their technical protocol in figures instead of narrate their methods only in words.

3. Material and methods part is baddly organized.

Reviewer #2: The rationale and objectives should be presented in separate paragraphs with titles. The abstract lacks a section on results, the format is truncated.

Reference 5 is not a trimera mouse model, should be removed as it does not pertain to the current subject.

Description of the humanized mice should include a review on the models available for HBV research.

Page 3 the authors state « The currently existing in vitro and in vivo HBV models make it possible to observe … through bioluminescence and fluorescence… allowing to assess the persistence and replication of viral DNA… do not require periodic euthanasia of the animal. » what is the litterature showing this? none to my knowledge, please cite the litterature.

Study methods section: « d) q » what does this mean?

What is the rational for irradiating mice after the grafts?

Page 8 the authors state « HBsAg in vivo visualisation is performed on Vilber Newton 7.0 … is used as a delivery agent » . what is the litterature showing this? none to my knowledge, please cite the litterature. The cited reference 41 only has in vitro data.

Animal ethics need to be better described and implemented: mice that undergo surgical procedures need to be treated with analgesic, not only anaesthetic, and humane endpoints of the maximum tumor size limits and weight loss should be included in the protocol description.

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Reviewer #1: No

Reviewer #2: No

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Protocol for chronic hepatitis B virus infection mouse model development by patient-derived orthotopic xenografts

PONE-D-21-25364R1

Dear Dr. Nagornykh,

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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Kind regards,

Md. Golzar Hossain, Ph.D.

Academic Editor

PLOS ONE

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