PONE-D-24-12332Vitamin K2 sensitizes the efficacy of venetoclax in acute myeloid leukemia by targeting the NOXA-MCL-1 pathwayPLOS ONE

Dear Dr. Takano,

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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: Yes

Reviewer #2: Yes

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

Reviewer #1: Yes

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: Yes

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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: Yes

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: Hematologists are increasingly faced with the appearance of resistance to the combined azacitidine - venetoclax therapy administered to patients with AML. That is why this article is welcome and its results deserve to be popularized, even if the number of included patients is small. It is a research direction for the future, which must be encouraged.

The authors described the research methodology very clearly, presented the results well and adequately discussed the pathophysiological mechanisms involved, including in the light of data from recent literature.

It is good for the authors to also discuss the effects of vitamin K on normal myeloid and erythroid progenitors: promoting the differentiation of myeloid progenitors and an anti-apoptotic effect that seemed to be dominant in erythroid progenitors (Sada E, Abe Y, Ohba R, Tachikawa Y, Nagasawa E, Shiratsuchi M, Takayanagi R. Vitamin K2 modulates differentiation and apoptosis of both myeloid and erythroid lineages. Eur J Haematol. 2010 Dec;85(6):538-48. doi: 10.1111/j.1600-0609.2010.01530.x. PMID: 20887388).

The authors must emphasize the fact that, according to the latest research, venetoclax resistance of AML cells involves transcriptional and phenotypic heterogeneity (Mohanty V, Baran N, Huang Y, Ramage CL, Cooper LM, He S, Iqbal R, Daher M; CTD2 Research Network; Tyner JW, Mills GB, Konopleva M, Chen K. Transcriptional and phenotypic heterogeneity underpinning venetoclax resistance in AML. bioRxiv [Preprint]. 2024 Jan 30:2024.01.27.577579. doi: 10.1101/2024.01.27.577579. PMID: 38352538; PMCID: PMC10862759).

It follows that extensive research is needed to investigate different potential therapeutic targets and lead to therapeutic solutions, probably combined or personalized.

The expression in English is very good.

In my opinion, the article can be published after minimal corrections.

Reviewer #2: This is a well-written manuscript and one of its kind as a proof of concept to encourage large-scale clinical trials. However, there are some significant gaps, such as why AZA is not considered alongside VK2 in the cell-based studies. This manuscript requires minor revision, as listed below:

1. Why AZA in combination with vk2 was not studied?

2. NOXA-MCL pathway explanation should be included in the introduction? It is a major pathway of the study and clearly took place in the title?

3. Cell line name should be provided appropriately, like SKM-1 – full name

4. Cell viability and apoptosis were performed on the knock down cell line or conventional? It was not mentioned in the methods – line 142, 150, 157, 166. Use the appropriate cell name in the methods.

5. What is FSC and SSC? (forward scatter and side scatter? of flow cytometry?)

6. Mention cycle 1 again in the result (line 253)

7. The extremely high response rate of what? (line 246). Provide the meaningful sentence

8. Is this the first study showing VK2-induced ROS damage the mitochondria ? (line 282)

9. Provide the acronym at first place as possible, ELN, ECOP

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Reviewer #1: No

Reviewer #2: No

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Vitamin K2 sensitizes the efficacy of venetoclax in acute myeloid leukemia by targeting the NOXA-MCL-1 pathway

PONE-D-24-12332R1

Dear Dr. Takano,

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.

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Kind regards,

Vivek Agrahari

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 #2: All comments have been addressed

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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 #2: Yes

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

Reviewer #2: 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 #2: Yes

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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

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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 #2: (No Response)

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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 #2: No

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