PONE-D-19-33856

Methylation of SRD5A2 promoter predicts a better outcome for castration-resistant prostate cancer patients undergoing androgen deprivation therapy

PLOS ONE

Dear Dr. Olumi:

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit and my decision is "Minor Revision" to fully meet  the 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.

Reviewer #1:

Understanding prostate cancer tumor progression mechanism and potential targets for treatment is very important. The 5a-Reductase types 1 and 2, encoded by SRD5A1 and SRD5A2, are key enzymes that catalyze the conversion of testosterone to dihydrotestosterone, androgen receptor (AR) agonist in prostate cells. 5a-Reductase type 2 is the predominant isoform expressed in the normal prostate. However, its expression decreases during prostate cancer (PCa) progression, whereas SRD5A1 increases, and the mechanism underlying this transcriptional regulatory switch is still unknown. In this research manuscript, Wang, et al  reports that methylation of SRD5A2 promoter occurs frequently in CRPC patients treated with ADT, and provided  important patient data (from local cohorts and TCGA). Furthermore, authors demonstrated the negative correlation between promoter methylation of SRD5A2 and SRD5A2 expression.

Comments:

1. The major strength of the study is the patient data from two cohorts to validate their hypothesis and comparison of findings with the TCGA data. Paper is well written and all the experiments with perfect controls in place.

2. Authors shall provide explanation about the basis on which CpG methylation sites were clustered into 3 modules, (CpG# -72 to -42; CpG# -40 to -35 and -31 to -19; CpG# -34 to -32 and -18 to 65 in CRPC patients) whereas methylation sites were clustered into 2 modules (CpG# -72 to -35, -31; CpG# -34 to -32 and -30 to 65 in metastatic CRPC cohorts).

3. Have the authors found any relation between the expression of SRD5A2 and survival in local CRPC patient cohorts, although they have shown through the TCGA data analyzed by MEXPRESS?

4. In Figure 3, 4 and 5 only 3 H has a value of “r”=0.78 (positive correlation), in other panels, “r” shows weak correlation, would the authors explain why.

5. In Figure 5 B and C, “r” and “P” values are not shown. Authors shall add the “r” and “P” in figure 5 B and C.

6. Have authors found any correlation between Androgen Receptor expression and SRD5A2 expression in local patient cohorts?

7. Authors have used local patent cohorts, please provide a link where data can be accessible if possible as per the PLosOne data sharing policy.

Reviewer #2:

The present study is a provacative analysis on the methylation status of the SDR5A2 promoter, which encodes an enzyme that metabolizes testosterone to the more potent androgen DHT, in castrate-resistant prostate cancer (CRPC)( specimens. Although the numbers of specimens analyzed in the present data are relatively small, the findings do show that there is a significant trend for promoter hypermethylation in CRPC specimens compared to benign prostate tissue. Most significantly, the higher ratio of promoter hypermethylation was positively associated with a significant increase in time to progression and patient survival when the patient data was interrogated. Immunohistochemistry analysis of SDR5A2 protein levels inversely correlated with the gene methylation levels, although this data has marked overlap and requires further follow-up. The data were further studied in the TCGA database which supported the primary data in this manuscript. Although the number of metastatic samples was very low in this study (n=12), the data indicate that the SDR5A2 methylation status is likewise able to predict patient survival.  The data are well discussed and fit well into emerging findings regarding the altered hormonal milieu in CRPC that drives androgen resistance.

Comments:

1. The paper would be improved by cell-based interrogations that replicate the specimen-based observations and support the conclusions, i.e. modifications of SDR5A2 methylation status change expression of this gene and alter its protein levels as well as steroid metabolism.

2. At present, the studies are merely correlative and direct studies are warranted. 

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University of Minnesota

Academic Editor, PLOS ONE

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(1) https://www.mdpi.com/2073-4425/9/9/429/htm

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(3) https://pubs.acs.org/doi/10.1021/acs.molpharmaceut.7b00070

(4) https://onlinelibrary.wiley.com/doi/full/10.1002/cncr.30071

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Methylation of SRD5A2 promoter predicts a better outcome for castration-resistant prostate cancer patients undergoing androgen deprivation therapy

PONE-D-19-33856R1

Dear Dr. Olumi,

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With Kind Regards,

Mohammad Saleem, Academic Editor-PLOS ONE

University of Minnesota-Minneapolis