PONE-D-22-26107Different androgen receptor-mediated pathways modulate cofilin phospho-regulation to control polarized migration of triple negative breast cancer lines of different molecular subtypesPLOS ONE

Dear Dr. Tahtamouni,

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. As indicated below, the reviewers have different opinions. As indicated by my comments, while there is a substantial body of data, it does not support the conclusions reached that coffin mediates androgen receptor activity in the TNBC cells studied. The manuscript is not therefore acceptable for publication in PLOS One. You are invited to address the reviewer comments and submit a revision but please note that all conclusions must be supported by the data provided. 

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Academic Editor

PLOS ONE

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Additional Editor Comments :

This is a comprehensive analysis of the effect of androgen stimulation on the migratory phenotype of three TNBC cell lines, and specifically its effect on cofilin modulation of the actin cytoskeleton. In general, there is a large amount of data that, while done appropriately, could be more clearly presented. Of major concern is the fact that the data does not support the conclusion of the authors that :

"Different androgen receptor-mediated pathways modulate cofilin phospho-regulation to control polarized migration of triple negative breast cancer lines of different molecular subtypes"

"Suppression of cofilin in MDA and to a lesser extend BT cells abrogated the androgen effects suggesting cofilin regulation is involved."

The only data that I was able to identify that supported a role for cofilin downstream of androgen receptor activation was changes in the N-cadherin/E-cadherin ratio that weren't followed up on in the paper. Most other effects of AR activation were cofilin independent and suggest that AR uses a cofilin-independent pathway to promote migration, as indicated at numerous places in the text by the authors. Inpaticualr, the data provided do not support the summary figure 10. As such, conclusions reached that cofilin mediates AR effects on TNBC cell migratory phenotypes are not supported by the data and the manuscript as such is not acceptable at PLOS One. Conclusions reached for a revised submission must be supported by the data provided and reflect the data obtained. I also suggest that the authors review all the data and present the data in a more concise and easy to follow manner that supports the conclusions drawn.

Comments:

1. Use of abbreviations for the cell lines used in the text and figures is not appropriate and full names of the cell line names must be used at all times.

2. Inclusion of fluorescent images without quantification is not acceptable (Fig 5, S4). Where quantified clear images should be provided showing the effect measured. For instance images of the AR effect on p-cofilin at the leading edge are not shown in figure 4.

3. The rational to include data in Table format is not clear and the extensive data provided in the tables should be presented as graphs.

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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: Partly

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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: No

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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: The readability and scientific structure/format of this manuscript is at least above average. It was not difficult to read/understand and many of the concepts were explained clearly and I did not find this review assignment any more difficult than if it was from another journal.

The manuscript and its findings used methods that were technically sound and used by many other cancer cell biologists in the field. The main value of this manuscript is for investigators in the field that are also looking at levels of very specific proteins (AR +/- DHT, p-cofilin) in the breast cancer cell lines chosen for analysis. In my opinion, its these kinds of reports that provide independent sources of external validation beyond meaning. I contend that there is no significant correlation between cofilin (p or not) and DHT activation amongst these cell lines but I do see value in these results being used to support other investigator's pursuits if they so happen to be looking for "...a paper that looked at the levels of p-cofilin with DHT activation and its effects on micro-actin tubules or invadopodia...".

Reviewer #2: The current manuscript investigates the potential relationship between AR stimulation and cofilin activity. Through AR stimulation and cofilin knock downs, the authors have assessed the role of AR in cofilin modulation and migration. The status and level of expression of other motility regulators have been evaluated.

Although there is a large body of work presented, there is no clear or specific connection demonstrated between AR and cofilin modulation. The observations are in fact expected for the most part. AR stimulates motility in the TNBC lines, therefore cytoskeletal reorganization such as cofilin dephosphorylation is expected. The same is true for proliferation. A cofilin KD would impair cell division. This is not specific to AR or any other growth or migration promoting agent.

There is also no data supporting the model proposed as it has not been investigated in this manuscript. The manuscript would also benefit from editing as it is too long and reads like a thesis.

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Reviewer #1: Yes: Hon Sing Leong

Reviewer #2: No

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The role of activated androgen receptor in cofilin phospho-regulation depends on the molecular subtype of TNBC cell line and actin assembly dynamics

PONE-D-22-26107R1

Dear Dr. Tahtamouni,

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,

Ivan R. Nabi, Ph.D.

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

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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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