Cisplatin and Starvation Differently Sensitize Autophagy in Renal Carcinoma: A Potential Therapeutic Pathway to Target Variegated Drugs Resistant Cancerous Cells

Round 1

Reviewer 1 Report

The manuscript titled "Cisplatin and starvation differently sensitize autophagy in renal carcinoma: a potential therapeutic pathway to target variegated drug-resistant cancerous cells" by Ankita Dutta et al. presents a study that explores the benefits of a starved condition for cisplatin treatment in renal cell carcinomas. This is an exciting topic because cisplatin treatment resistance or failure is still challenging in clinical treatment. However, there are some questions that need to be addressed regarding the study.

1.     In Figure 1, demonstrating their hypothesis with only one cancer cell line may not be sufficient.

2.     The authors indicate that nutrient deprivation effectively triggers autophagy, as evidenced in Figure 1A. Nonetheless, it remains to be verified whether the enhanced cytotoxicity observed in cisplatin treatment during starvation is linked to autophagy, as illustrated in Figure 1D. In order to establish this correlation, it is recommended to employ knockdown autophagy-associated genes.

3.     I had trouble reading some of the words in the figures. Specifically, I found Fig2C-E, Fig2F(d), Fig3B(a,b), Fig3C(b), and Fig3D(b) to be unclear.

4.     The authors utilized the Autophagic ACHN cell line and autophagic ACHN cell lines treated with cisplatin to conduct mRNA-seq and pinpoint hub genes. However, in Figure 5A, they confirmed these hub genes by employing qRT-PCR on the ACHN cell line under two distinct conditions: Control (untreated cells) and PBS (cells deprived of nutrients for three hours). Why? Please explain it.

5.     Please note that in line 28, the abbreviation Qrt-PCR should be corrected to either qRT-PCR or RT-qPCR.

6.     The sentence in line 485 needs to be revised. It currently reads, "under nutrient nutrient-deficient environment…", but one "nutrient" should be removed for clarity.

Minor editing of English language required

Author Response

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Reviewer 2 Report

The subject addressed in this article is interesting because there is a huge unmet need to find reverse cisplatin resistance in cancers. I suggest Acceptance of current MS with minor revision keeping in consideration the comments given below:

Suggestions for authors:

       I.            As autophagy is the main mechanism so authors need to provide strong supportive proofs for the same at least LC3 I to II conversion and P62 degradation.

    II.            To further support the central role of autophagy, authors can further support their results using autophagy modulators or genetic manipulations.

 III.            Though authors have provide mRNA results but for apoptosis, authors need to provide protein levels change of apoptosis indicators.

 IV.            Fluorescence images also need to be replaced with images of publishable quality.

    V.            In method part, Authors need to mention the fluorescence filters for both annexin and PI to make it repeatable by scientific community.

English is up to the mark

Author Response

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Reviewer 3 Report

Cisplatin has been a mainstay of cancer chemotherapy since the 1970s. Despite its broad anticancer potential, its clinical use has regularly been constrained by kidney toxicities. The drug was first approved as an antineoplastic agent in 1978 and remains an important and effective therapy today for the treatment of various cancers including bladder, breast, cervical, esophageal, head and neck, ovarian, prostate, small and non-small cell lung, stomach, testicular cancers, Hodgkin’s and non-Hodgkin’s lymphomas, melanoma, mesothelioma, multiple myeloma, neuroblastoma, and sarcoma. 

I think the direction to choose cisplatin as a chemotherapeutic agent in renal carcinoma is not proper. Additionaly, cisplatin and its mechanisms of action are broadly tested. High nephrotoxicity is a reason to find the analogues of cisplatin with better toxicological profile.

The conclusion of the paper sounds unprofessional: "This study will pave the way for clinical studies to begin for better drug efficacy during chemotherapy while on a restricted diet or fasting.

I don't see the high novelty and originality of this article. I decided to reject the paper.

Author Response

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Round 2

Reviewer 1 Report

Thank you for your response. However, some pathways in Figure 3B(a) were covered by the spot and need to be revised.

Author Response

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