Stress Granule-Driven Resistance in Cancer: Mechanisms and Emerging Strategies

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

1. A brief section on components of stress granules can be a great asset of this review.
2. Section 2: This section explains how stress granules assemble and dissemble, but it does not clarify whether these dynamic processes influence stress granule driven resistance in cancer. Since stress granule remodeling is known to affect how tumor cells cope with therapy-induced stress, adding a brief explanation of this link would improve the relevance of the discussion.
3. The manuscript would benefit from briefly distinguishing SG functions in normal cells versus cancer cells, as SGs in tumors show altered composition and persistence that drive therapy resistance.
4. The authors describe several SG-related mechanisms, but a brief and unified tabular summary of the key signaling pathways driving SG formation and function in cancer is missing.
5. The manuscript could be strengthened by discussing methods to quantify SGs, such as microscopy based puncta counts, live cell imaging, FRAP, or biochemical enrichment and correlating SG abundance or dynamics with therapy resistance in clinical samples.
6. The manuscript discusses SG induction by anticancer drugs and upregulation of SG proteins in cancer, but it does not specifically summarize which signaling pathways are targeted by small molecules or compounds to inhibit SG formation. Including a concise overview of these pathways and corresponding compounds would strengthen the clinical relevance and provide clearer guidance for therapeutic strategies.
7. A figure summarizing SG-mediated drug resistance in cancer cells would greatly enhance the manuscript.

Comments on the Quality of English Language

Syntax part should be considered.

Author Response

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Reviewer 2 Report

Comments and Suggestions for Authors

This wonderful review article arrives in a timely manner, which was much needed in the field. It is very well written, with therapeutic implications clearly highlighted, and the illustrative figures are excellent in conveying complex concepts. The manuscript provides a comprehensive overview of stress granules (SGs) in cancer biology and therapy resistance. However, before acceptance, several major revisions are required to strengthen clarity, mechanistic depth, and translational relevance:

 

Major Comments

1. Clarify Mechanistic Pathways and Distinguish Canonical vs. Non-Canonical SGs The manuscript describes both canonical and non-canonical SG assembly, but the distinctions are sometimes blurred. Authors should provide a clearer comparative framework (e.g., a summary table or schematic) that explicitly contrasts canonical ISR-driven SGs with non-canonical SGs induced by drugs or alternative pathways. This will help readers better understand the diversity of SG biology in cancer.

2. Expand on Tumor Microenvironment and Clinical Implications While the tumor microenvironment is mentioned as a driver of SG formation, the clinical implications are not fully integrated. Authors should expand on how hypoxia, nutrient deprivation, and therapy-induced stress specifically shape SG dynamics in vivo, and how this knowledge could inform rational combination therapies. A dedicated subsection summarizing translational opportunities would strengthen the therapeutic message.

3. Address Methodological Limitations and Research Gaps The manuscript should more explicitly discuss the challenges of studying SGs in cancer, such as reliance on in vitro models, variability in SG markers, and difficulties in visualizing SGs in patient tissues. Proposing methodological solutions (e.g., advanced live-cell imaging, single-cell transcriptomics, or proteomic approaches) would add depth and highlight future research directions.
   

   4. Oncogenic RAS and Stress Granules in Therapeutic Contexts It has been shown elegantly that oncogenic RAS influences stress granules in therapeutic aspects and reflects the latest understanding of SGs and their role in tumor progression and therapy resistance (PMID: 31911550, 37179344). Authors must add a few lines discussing this aspect with the cited references, as it directly connects oncogenic signaling, SG dynamics, and therapy resistance. This addition will broaden the scope of the review and link SG biology to oncogene-driven cancer therapy.

 

Summary

This is a strong and timely manuscript with excellent writing and figures. With the above revisions — (1) clearer mechanistic distinctions, (2) deeper integration of tumor microenvironment and clinical implications, (3) acknowledgment of methodological limitations, and (4) inclusion of oncogenic RAS influence on SGs — the paper will be significantly strengthened and ready for acceptance.

Author Response

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Author Response File: Author Response.docx

Reviewer 3 Report

Comments and Suggestions for Authors

1. Abstract: After line 28/29, the authors should state the objective of writing this review.
2. In the introduction, several anticancer drugs are mentioned. Discuss previous studies of those anticancer drugs.
3. The presence of SGs plays a major role in cancer cells' adaptability to the tumor microenvironment. Authors should briefly discuss the TME and the adaptability. 
4. Briefly explain why SGs are important in cancer, identify the existing gaps in SG cancer research, and clarify how this review provides new insights or advances the field.
5. There are unnecessary hyphens in many places; remove them.
6. Canonical vs. non-canonical SGs are described, but add 1–2 sentences on how each type influences cancer progression.
7. In Section 3, include stress levels or drug concentrations (e.g, arsenite dose, lapatinib dose)
8. Most of the references are old. Cite recent studies.
9. In Section 4, some mechanisms are described without stating the experimental model (cell line/animal/patient samples).
10. Section 4.3: Authors are advised to discuss the ER stress in a better manner by following and citing the 10.1080/10717544.2023.2284684
11. Several short paragraphs should be combined to improve flow.
12. In the drug-induced SG section, consider grouping drugs into those categories (kinase inhibitors, DNA-damaging agents, proteasome inhibitors, etc.)
13. Provide more detail on how these stress-responsive kinases act differently in cancer vs. normal cells.
14. At the beginning of the section on SG protein upregulation, add a short explanation of why this overexpression matters in cancer biology.
15. Authors can add 1–2 sentences explaining how these PTMs promote SG localization, particularly for NEDDylated SRSF3, ribosomal proteins, and Cullin
16. When referring to “specific kinases,” please list the four ISR kinases (PERK, PKR, HRI, GCN2).
17. The tables repeat the same studies which is already in the text. Add new information to tables to provide readers with more data.
18. Some sections heavily focus on mechanisms; adding more clinical relevance can improve this review.
19. Please check for grammar and punctuation consistency, especially long sentences that can be split for clarity.

Comments on the Quality of English Language

Many grammatical errors, and there are many long and complex sentences. Authors are advised to correct those. 

Author Response

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Reviewer 4 Report

Comments and Suggestions for Authors

Review Report

Concerning the manuscript no. cancers-4015188

Entitled „Stress Granule–Driven Resistance in Cancer: Mechanisms and Emerging Strategies:

 

General opinion: The manuscript presents an up-to-date, well-documented, and logically organized review of the role of stress granules (SGs) in cancer resistance to treatment and discusses potential therapeutic strategies targeting SGs. The text is coherent, written in clear language, and the literature is very current (numerous papers from 2023–2025). In my opinion, the strengths of this manuscript are: (i) the currency and breadth of the cited literature, (ii) the good structure and logic of the argument, (iii) the strong connection to clinical oncology, (iv) the clear delineation of challenges and knowledge gaps, particularly in Section 7 ("Challenges/Outstanding Questions").

 

 

Major comments:

1. In section 4, the authors extensively discuss chemotherapeutic drugs and kinase inhibitors, while the role of SGs in response to radiotherapy and immunotherapy is only marginally outlined (mainly through isolated mentions of YBX1 after radiation and possible immunomodulatory effects in the discussion). I suggest adding a short subsection in section 4 or 8 that would summarize what we know about SGs in the context of radiotherapy and how SGs may influence response to immunotherapy (checkpoint inhibitors, interferon modulation, RIG-I, dsRNA-sensing).
2. Sections 4.3 and 4.4 are very rich in content, but sometimes it reminds me a list of examples. A s for me, more concise conclusion to each of them would be helpful, clearly summarizing how the described phenomena translate into treatment resistance (e.g., modulation of proapoptotic mRNA availability, metabolic reorganization, alteration of immune signals).
3. The section on G3BP1/2 is very good and strong, but requires a somewhat clearer practical conclusion.
4. Section 6 provides a good description of the multifunctionality of G3BP1/2 (mTOR, mitophagy, insulin secretion, Wnt/β-catenin, viral infections). It would be worthwhile to add a section in the final paragraph of this section providing practical guidance to the clinical/therapeutic reader, noting that "pure" targeting of G3BP may require strategies to distinguish SG-dependent from SG-independent functions (e.g., selective modulation of the domains responsible for condensation rather than complete protein degradation), and that biomarkers monitoring the side effects of such targeting should include not only SGs but also, for example, mTOR activity and metabolic parameters.
5. Section 5 and Table 2 describe many interesting compounds (MLN4924, PARP inhibitors, G3Ia/G3Ib, C108, PROTAC, lipoamides, ISRIB). It would be useful to add another column to the table indicating whether a given compound is in preclinical development, in Phase I–II clinical trials, or is already an approved drug and is being considered for repurposing in the context of SG, etc.
6. The authors rightly emphasize the correlation of high expression of G3BP1/2, YB1, and IGF2BP3 with poorer prognosis. To make this section more practical, I suggest adding a short paragraph (e.g., in the discussion) summarizing whether and how these markers can be used as potential biomarkers predicting treatment response, markers for patient selection for SG-targeted therapies, or pharmacodynamic markers in clinical trials of SG-modulating molecules.

 

 

Minor comments:

1. Please ensure that all abbreviations are defined upon first use in the main text or in a table (e.g., PROTAC, EPS, IDPs, PDX, UPR, P-bodies).
2. In the text, it is advisable to refer directly to Tables 1 and 2 and Figures 1 and 2 more frequently when discussing the same topics (e.g., when describing SG-inducing drugs and condensate modulators). This will make it easier for the reader to quickly transition between the description and the graphical summary.
3. The captions under Figures 1 and 2 could be more descriptive (e.g., explicitly indicate which steps are dependent on eIF2α-P and which on eIF4F/4EBP1; which interventions are already being tested in vivo).

Author Response

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Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

1. While the authors have mentioned several SG quantification approaches, the revision largely lists method names without elaboration. To adequately address this point, a separate, dedicated subsection discussing each quantification method (principle, strengths, and limitations) would be appropriate, rather than brief mentions.

2. Although Table 3 is interesting, the clinical value of SG-targeting drugs is mostly unclear. A brief discussion of the translational importance of these approaches, rather than simply stating their developmental stage, would increase the review's quality and thoroughness.

Author Response

Comment 1: While the authors have mentioned several SG quantification approaches, the revision largely lists method names without elaboration. To adequately address this point, a separate, dedicated subsection discussing each quantification method (principle, strengths, and limitations) would be appropriate, rather than brief mentions.

Response 1: Thank you for emphasizing this point. We have now addressed this point in detail as follows: "Researchers typically visualize SGs using immunofluorescence or live-cell imaging of established marker proteins such as G3BP1, TIA-1, or eIF3 subunits. These approaches enable direct visualization of SGs at high spatial resolution and the assessment of cell-to-cell variability. Quantitative parameters include SG number per cell, size, total area, and subcellular distribution. Live-cell imaging further permits temporal analysis of SG assembly and disassembly dynamics in response to stress or pharmacological interventions. However, microscopy-based approaches may not readily distinguish bona fide SGs from other cytoplasmic ribonucleoprotein aggregates without careful marker validation. Fluorescence recovery after photobleaching (FRAP) assesses the dynamic properties of SGs by measuring the exchange rates of fluorescently labeled SG-associated proteins within granules. Recovery kinetics provide insights into SG material properties, including liquidity, protein mobility, and maturation state. This approach offers quantitative, kinetic information that static imaging cannot capture and is particularly useful for distinguishing liquid-like SGs from more solid or pathological assemblies. However, photobleaching itself may perturb SG integrity, and recovery measurements can vary depending on the selected marker protein and stress condition."

Comment 2: Although Table 3 is interesting, the clinical value of SG-targeting drugs is mostly unclear. A brief discussion of the translational importance of these approaches, rather than simply stating their developmental stage, would increase the review's quality and thoroughness.

Response 2: Thank you for the suggestion. We have added this point as follows: "Therefore, SGs represent a rapidly reversible, non-genetic survival strategy that cancer cells exploit to withstand chemotherapy, targeted inhibitors, and microenvironmental stress. Because SGs suppress apoptosis, preserve pro-survival transcripts, and buffer cells against proteotoxic and metabolic stress, therapeutically disrupting SG assembly or accelerating their clearance can convert transient drug tolerance into prolonged tumor control and delayed disease recurrence. Importantly, multiple SG-targeting strategies, including PTM inhibitors, biomolecular condensate modulators, stress-kinase blockers, and indirect metabolic interventions, have been shown to enhance the cytotoxicity of standard chemotherapeutics in preclinical models. These findings underscore a key translational principle: SG-directed agents improve the efficacy of existing therapies by disrupting the stress-adaptive mechanisms that enable cancer cells to survive treatment. As such, rational combination regimens that pair SG inhibition with chemotherapy, kinase inhibitors, or immunotherapy hold substantial promise for overcoming drug resistance and improving clinical outcomes."

Reviewer 2 Report

Comments and Suggestions for Authors

Although PMID: 31911550 was suggested in the earlier review, it has not been adequately discussed. This study clearly demonstrates a translational approach to targeting stress granules in KRAS-driven cancers by restricting glutamine metabolism, which in turn plays a role in stress granule regulation. In the absence of direct stress granule inhibitors, the use of glutamine metabolism inhibitors provides an important translational strategy to modulate stress granule dynamics. The authors should add a few lines discussing this aspect and appropriately cite the study.

Author Response

Comment 1: Although PMID: 31911550 was suggested in the earlier review, it has not been adequately discussed. This study clearly demonstrates a translational approach to targeting stress granules in KRAS-driven cancers by restricting glutamine metabolism, which in turn plays a role in stress granule regulation. In the absence of direct stress granule inhibitors, the use of glutamine metabolism inhibitors provides an important translational strategy to modulate stress granule dynamics. The authors should add a few lines discussing this aspect and appropriately cite the study.

Response 1: Thank you for explaining this insight. We have now cited the suggested manuscript and added a dedicated subsection (subsection 5.4) explaining indirect modulators of SGs, as follows: "SGs can also be targeted indirectly by modulating metabolic and stress-sensing pathways or by enhancing SG clearance. One such approach involves using glutaminase inhibitors to modulate SG assembly in KRAS-driven therapeutic resistance. Oncogenic KRAS upregulates NRF2, a master regulator of oxidative stress responses, thereby promoting metabolic rewiring and increased glutamine dependence, which contributes to resistance to gemcitabine treatment [90]. Glutamine deprivation or pharmacologic inhibition of glutaminase attenuates SG formation under these conditions, disrupts redox homeostasis, and sensitizes cancer cells to gemcitabine. Clinically approved autophagy activators, such as rapamycin or trehalose, can also enhance SG clearance; however, determining their effectiveness in modulating SGs and inhibiting therapy resistance remains a robust area of investigation."

Reviewer 3 Report

Comments and Suggestions for Authors

May be accepted for publication. 

Author Response

Comment: May be accepted for publication. 

Response: We thank the reviewer for this positive evaluation of our manuscript.