PONE-D-22-00370

A Pyridinesulfonamide Derivative FD268 Suppresses Cell Proliferation and Induces Apoptosis via Inhibiting PI3K Pathway in Acute Myeloid Leukemia

PLOS ONE

Dear Dr. Zhou,

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 by both Reviewers.

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Francesco Bertolini, MD, PhD

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: 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: Congratulations on the excellent article. Some considerations:

- in keywords: you do not need to include those that are already mentioned in the title (pyridinesulfonamide; acute myeloid leukemia; Apoptosis)

- objectives are not well highlighted

- in some tests, in the text of materials and methods, it is not clear whether triplicates and 3 independent experiments were carried out, or only 3 independent experiments. They are only explained in the subtitles.

- in figure 4c the name of the cell lines was missing

- the work has been done with two cell types, but in RNAseq only one cell line was chosen, but there was no explanation for this.

Reviewer #2: The manuscript “A Pyridinesulfonamide Derivative FD268 Suppresses Cell Proliferation and Induces Apoptosis via Inhibiting PI3K Pathway in Acute Myeloid Leukemia” by Chen et al. presents the results on therapeutic activity of novel and highly active FD268 inhibitor of PI3K. The authors found that FD268 inhibits the viabilities of AML cells with the efficacies superior to other two PI3K inhibitors, PI-103 and CAL-101. They showed that FD268 suppresses cell proliferation, induces cell cycle arrest and apoptosis of AML cells through the inhibition of PI3K activity.

In my humble opinion, I think that this work will be of interest to a broad audience.

Overall, the manuscript is well written and experiments were properly designed and executed. However, some minor corrections and improvements are necessary prior to its acceptance.

Comments and Suggestions:

Line 90: repeating words “and and”.

Line 108: Should read “…and treated with FD268…”

Line 178: Should read “…plates and treated with FD268 (0.125, 0.5 µM) or DMSO for 24 h. Cells were harvested…”

Line 225: Should read “The values were shown as the…”

Lines 249-250: Fig. 3C: there is no dose dependent inhibition of cleavage of caspase 3 and PARP, but increase instead. The sentence should be corrected as “…proteins were consequently examined by western blot assay, which showed a dose dependent increase of both caspase-3 and PARP cleavage (Fig 3C).”.

Line 258: Should read “Statistical significance is represented with asterisks *P < 0.05 and ***P < 0.001.”, as there is no **P < 0.01. It might be also better to use white background for S phase data, as labels/asterisks are barely visible in Figure 2B.

Figure 4C: Indicate which blots represent HL-60 and MOLM-16 cells. Label for cell lines is missing.

Line 288: The authors should mention here that the protein levels of other markers of apoptosis, e.g. Bcl-2, Bax and Bcl-XL, did not change significantly in FD268-treated cells.

Figure 5C: Label of MOLM-16 has to be corrected (swapped letters). The same applies to Figure 6A and 6B.

Figure 9: Shouldn´t the cleaved caspase-3 and PARP lead to cell death/apoptosis, instead of cell survival (check the bottom right corner)?

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Reviewer #1: No

Reviewer #2: No

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PONE-D-22-00370R1A Pyridinesulfonamide Derivative FD268 Suppresses Cell Proliferation and Induces Apoptosis via Inhibiting PI3K Pathway in Acute Myeloid LeukemiaPLOS ONE

Dear Dr. Zhou,

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.

==============================

ACADEMIC EDITOR: Please insert comments here and delete this placeholder text when finished. Be sure to:

• Indicate which changes you require for acceptance versus which changes you recommend
• Address any conflicts between the reviews so that it's clear which advice the authors should follow
• Provide specific feedback from your evaluation of the manuscript

Please ensure that your decision is justified on PLOS ONE’s publication criteria and not, for example, on novelty or perceived impact.

For Lab, Study and Registered Report Protocols: These article types are not expected to include results but may include pilot data. 

==============================

Please submit your revised manuscript by Nov 19 2022 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,

Jianhong Zhou

Support Staff - Editorial

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 (if provided):

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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

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 #1: Yes

Reviewer #2: Yes

**********

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

Reviewer #1: Yes

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 #1: Yes

Reviewer #2: 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

Reviewer #2: 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: Now, The manuscript is much better! All notes have been answered.

The reading is better understood after all mistakes are solved.

Reviewer #2: The authors have revised the manuscript according to my suggestions and recommendations. I have 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: 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.

A Pyridinesulfonamide Derivative FD268 Suppresses Cell Proliferation and Induces Apoptosis via Inhibiting PI3K Pathway in Acute Myeloid Leukemia

PONE-D-22-00370R2

Dear Dr. Zhou,

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,

Jianhong Zhou

Staff Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments: