Review Reports - The Crucial Role of AR-V7 in Enzalutamide-Resistance of Castration-Resistant Prostate Cancer

Round 1

Reviewer 1 Report

Dear Authors,

I reviewed with interest the paper entitled “The crucial role of AR-V7 in Enzalutamide-resistance of Castration-Resistant Prostate Cancer”.

First, I would strongly congratulate with the authors for their hard work for the present study, which covers an actual and very interesting topic such as mechanisms of resistance of systemic therapies in CRPC patients. Indeed, in this study authors aimed to summarize the mechanisms and biological behaviors of AR-V7 in Enzalutamide of CRPC.

I found the present study interesting and well written - no major concerns with language editing or general fluency.

However, before further evaluating the article, I would recommend revision to be taken to improve the quality of the manuscript - below my specific comments:

- The title is clear and descriptive of what authors have explored in their work, yet my suggestion is to include also the term a term for the specific type of article (e.g., “review”).

- The Introduction provides a background which is relevant to the study, yet it could be further improved. Indeed, I would recommend mention that Enzalutamide is a treatment option in both non metastatic and metastatic CRPC patients, with proper references (e.g., DOI: 10.3389/fonc.2021.700258; DOI: 10.1016/j.eururo.2016.08.002). With specific regards to AR-V7, to underline its pivotal role in the natural history of PC, and for completeness of the topic, I would recommend mentioning also its role - which has been recently evaluated - in predicting risk of progression in the setting of early-stage PC (DOI: 10.1159/000512445).

- The aim of the study is properly stated at the end of the paragraph.

- Figures are clear and not repetitive, as well as all the sub-headings of the manuscript.

- Overall, the paper results methodologically correct and well written. However, I would recommend adding a paragraph specifically reporting the current limitation of data present in the literature on this topic.

I have not further suggestions.

Author Response

Dear Reviewers:

Thank you for your letter and for the reviewers’ comments concerning our manuscript entitled “The crucial role of AR-V7 in Enzalutamide-resistance of Cas-tration-Resistant Prostate Cancer” (ID:cancers-1918727).

Those comments are all valuable and very helpful for revising and improving our paper, as well as the important guiding significance to our researches. We have studied comments carefully and have made corrections which we hope to meet with approval.

Please see the attachment.

Sincerely

Chen Shao

Author Response File: Author Response.pdf

Reviewer 2 Report

Figure1

Please correct from HR to HD.

Heat shock protein:

・Authors should explain where and how the HSPs combine and work in the AR/AR-V7.

It is difficult to understand why anti-HSP drugs restore sensitivity for ENZ.

Author Response

Dear Reviewers:

Please see the attachment.

Sincerely

Chen Shao

Author Response File: Author Response.pdf

Reviewer 3 Report

Revision

Revision of manuscript ID 1918727 titled “The crucial role of AR-V7 in Enzalutamide-resistance of Castration-Resistant Prostate Cancer” by Zheng Z. et al, to Cancers.

In this review the authors focus on the molecular mechanisms controlling the protein stability of the AR-V7 as well as the effect of LncRNAs on pathways and cell signaling that modulates AR-V7. The overall idea is to understand the complex mechanisms that control AR-V7 expression and how to tackle them in order to improve and design new strategies to overcome Enzalutamide resistance in mCRPC patients.

Overall, the idea is good, although it is very focus on only one specific treatment approved for patients with CRPC and on the proteostasis of AR-V7 and some general mechanisms controlled by LncRNAs. The manuscript can be followed, although I found many sentences that need to be reviewed as they are hard to follow, or they are not following the context of the paragraph, or they are just empty statements that do no help or conclude. There also some carelessness mistakes. I encourage the authors to address the comments.

Major comments

1. Although the focus in AR-V7, I strongly suggest the authors to include in the Figure 1, all the other AR variants that were mentioned in Section 2. Otherwise, it is really hard to follow the differences between them by just reading.

2. The authors should modify Table 1, to include a column discussing if any of the compounds mentioned are at preclinical or clinical stages, or even approved. That will help readers. I also recommend that the column named as “mechanism” to be divided in to two columns one to determine the target(s) of the compound mentioned and the second column to summarizes the regulatory mechanisms.

3. I strongly think that sections 4.1.2 and 4.1.3 should be moved out to section 4.3. Also, modify the subheading to make it more related to the present study.

4. Table 2 heading “Effect of CRPC on ENZ sensitivity” should be modified to better describe the column content. This make not much sense as you are not looking into the effect of CRPC, you are determining the effects of specific genes in ENZ sensitivity. Also, the sensitivity is not inhibited! Please define better the content of this Table!

5. In Table 3, in the second column the authors described “Sample number (CRPC/Control). What is the definition of Sample number? Are they discussing patients, cell lines, tissues, or what? Please describe properly. Also, add a column to indicate the type of sample assessed in each study. That will be of much help for readers. Column 4 should be better explained as inhibit/promote are not related to ENZ sensitivity.

6. In Table 3, please modify the title to make it clearer: “Expression of LncRNA in CRPC and its sensitivity to ENZ.” Also, unify the Up and UP!

7. Sentence I believe the authors should consider to re-write s they are really hard to follow:

a. Lines 45-50: “Androgen deprivation therapy (ADT) has become the standard treatment of PCa owing to such a close correlation between androgens and PCa. It helps the reduction of the androgen level or the inhibition of androgen receptor (AR) in the human body through surgery or drug castration such as the application of the AR signaling inhibitor (ARSI). As a result, it functions as the treatment or inhibits the progression of PCa.”

b. Lines 54-55: “It was approved to be 54 listed in China at the end of 2019 for the treatment of CRPC.”

c. Lines 79-81: “It is in the field of PCa, in particular, that the change of AR-VS proteostasis and its relations with LncRNA has attracted the attention of researchers with thereby in-depth research conducted.”

d. Lines 317-320: “Meanwhile, LncRNA can regulate its transcription level and epigenetic modification through interaction with DNA, proteostasis, and so forth [102, 103].

e. Lines 343-346: “Meanwhile, LncRNA-Malat1 is indispensable for the ENZ-induced expression of AR-V7, as proved by the fact that using siRNA for the knockdown of Malat1 to inhibit its expression resulted in a marked decrease in AR-V7 expression.

f. Lines 384-386: “AR-V7 is the most pertinent AR-VS that has been confirmed to the resistance mechanism and is crucial in mediating CRPC resistance to the existing medication and predicting therapeutic response.”

g. Lines 392-394: “For example, the conventional drug niclosamide has been applied in a new field. There have been also ARVib-7 and MTX-23, newly developed small molecule compounds, and natural products, nobiletin and Hsp90.”

Minor comments

1. Make sure all gene names utilized in the review are italicized and follow proper nomenclature.

2. In the legend of Figure 2, you need to define AR-FL and AR-V7.

3. Please make sure you abbreviate the words the first time you mentioned. For example: ENZR. It was defined in the Abstract but not in the main text.

4. In the abstract, please review this sentence: “However, almost all patients performed drug resistance after PCa during AR antagonist treatment because of various mechanisms.” Patients do not perform drug resistance! Please correct the terminology.

5. Is this the correct format for the number “380 thousand”? please check the guidelines.

6. Please be consistent with nomenclature of LncRNAs. Refer to Line 333 LncRNA-Malat1 or Table 3 LncRNA-MALAT-1. Unify!

7. Typos/errors:

a. Line 392: “the AR-391 V7 protein or the AR-V7 transcription level.” Levels!

b. Lines 367-368: “In 367 sum , targeting”. Extra space!

Author Response

Dear Reviewers:

Please see the attachment.

Sincerely

Chen Shao

Author Response File: Author Response.pdf

Round 2

Reviewer 3 Report

I want to thanks the authors for the effort in addressing all my comments in such a short time. The review has significantly improved, but still needs some editing. I suggest the authors to carefully review the manuscript as in the rush of correcting and having it out there are multiple typos and errors.

Just to point out some:

Please refer to Lines 38: "...in2020."

Please refer to Lines 45: "...PCa.Consequently ,"

Please refer to Lines 55: "non Mcrpc"

CPRC instead of CRPC in lines 57,373, 375.

Please refer to Lines 85: subunit 1(MED1)[29], forkhead box01 (FOX01)[30]、GATA

In Table 2. I suggest the authors to please review the term "patients" and changed it for either tissues or cell lines, if these sample types were analyzed in Neha Singh et al (2021) [Ref 134].

This sentence is still confusing: "By the observation of the single splicing connection in cells and tissues can this technique overcome the limitation that AR-V7 are not able to be detected precisely [67].

This sentence needs to be review: "...and consequently made progress for efficient detection efficiency [68].

The heading in Table 3 is shift and not center!

Remove number 1 from line 880.

Author Response

Dear Reviewers:

Thank you for your comments concerning our manuscript entitled “The crucial role of AR-V7 in Enzalutamide-resistance of Castration-Resistant Prostate Cancer”(ID:cancers-1918727).

Those comments are all valuable and very helpful for revising and improving our paper, as well as theimportant guiding significance to our researches. We have studied comments carefully and have made corrections which we hope to meet with approval.

We tried our best to improve the manuscript, we appreciate for Editors/Reviewers’ warm work earnestly,and hope that the correction will meet with approval. Once again, thank you very much for your comments and suggestions.

Sincerely

Chen Shao

Author Response File: Author Response.docx