Development of Olaparib-Resistance Prostate Cancer Cell Lines to Identify Mechanisms Associated with Acquired Resistance
Round 1
Reviewer 1 Report
The authors review mechanisms for development of clonal resistance in PARP inhibition in prostate cancer. This research is novel in that the authors correctly identify mechanisms of drug resistance in prostate cancer are less well studied. They provide a comprehensive analysis for this foundational work. I believe the research fits with the journal mandate and is appropriately conducted. I have the following comments. I very much enjoyed reading this paper overall it is well designed and conducted study. The manuscript is well written and I feel it fits within the mandate of Cancers.
Abstract: No specific concerns.
Introduction: The authors provide sufficient background information for and detail the clinical context for this study.
Methodology: The experimental design is appropriate for this type of analysis. The authors provide sufficient detail such that we would be able to replicate the experiment.
Results:
It is interesting that cell doubling time was not different between OR and WT cell lines. I naively would have expected a post-therapy accelerated repopulation to occur. Can the authors comment on this / explain in the discussion for those of us with more clinically oriented understandings of cell behavior?
The authors should comment specifically on why ROCK2 upregulation might be seen in all OR lines.
Discussion:
I would like to see the authors expand the discussion as it does appear abbreviated. They do discuss their results in the context of breast cancer and parp inhibitor resistance but do not provide more granular detail on several key findings within their study. For example, can they comment on the above. Otherwise, can they comment on potential pathway exploitations that may show benefit in combination therapies for OR cancers? Is there a plan for subsequent study of this? There are many weaknesses of this type in vitro study. The authors should be cognizant of these and be open with their readers in explaining the same.
Author Response
Please see the attachment.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Congratulations on addressing an important topic such as pathways involved in Olaparib resistance in PC. I really appreciated the methodology. The manuscript is suitable for publication after a minor revision
- you discussed autophagy as a pivotal element in chemotherapy resistance, however, you should underline how complex the process is. Indeed, depending on the cellular context, autophagy could play either a detrimental or a protective role in PCa survival due to its complex modulation and pleiotropism. I recommend discussing this important topic taking into consideration the most recent finding on autophagy and PCa. You can find an interesting point in this article which I recommend including in the references (PMID: 35409187 PMCID: PMC8999129 DOI: 10.3390/ijms23073826)
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Reviewer 3 Report
Cahuzac and co-authors conducted a study entitled “Development of olaparib-resistance prostate cancer cell lines to identify mechanisms associated with acquired resistance”, in which they elucidating the development of multidrug resistance as a result of treatment of liver cancer patients with transcatheter chemoembolization (TACE). They applied a qRT-PCR technique to determine the change in MDR-related microRNAs levels using blood from patients and liver cancer cells HepG2 resistant to doxorubicin. The authors reported a decrease in miR-223-3p level in TACE-resistant patients. On cellular level, miR-223-3p targeted MDR expression. Although the current study is well-designed and provided invaluable information; however, I have few comments to improve the study.
- In the abstract section, please define HR (line 29) and replace prostate cancer (line 31) with PC. In the keywords, please add inhibitor after PARP “PARP inhibitor”.
- In line 49, please define HRR and NHT. Line 53, replace prostate cancer by PC. Reference(s) needs to be included to support the statement in line 57-57.
- Line 90, please define STR.
- Have the authors looked at any genes linked to resistance and AR expression in these 3 cells?
- What was the rational on which the authors selected the three candidate genes?
- I suggest to either keep Fig. 1b or Table 1, but we cannot keep both. In figure 1 legend. Please add a statement of data are statistically significance at p<0.05
- In results section, what is the rational to use the cut-off value for up and downregulated genes and Table 4 needs more explanations. Why BRCC3 recognized in Table 4 and not recognized in Table 3?
- In Fig. 3 legend, please removed *** P<0.001 because it is not included in the figure.
- In Fig. 4, B & C, be consistent in reporting the statistical significance and remove “and” from the legend.
- Please remove Fig. 3b from the discussion.
- To assure that these genes are associated with OR, the authors should at least knockdown one gene as a proof of concept in resistant cells. If the cells become sensitive, it tells the gene is involved in OR.
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
The authors have addressed my comments.
Reviewer 3 Report
The authors addressed most of the raised concerns.
