Functional Assays to Guide Personalized Oncological Treatment of Patients with Soft-Tissue Sarcomas
Round 1
Reviewer 1 Report
Comments and Suggestions for Authors
The article Functional assays to guide personalized oncological treatment of patients with soft-tissue sarcomas.is an interesting review on the direct tests of the drugs on cells or pieces derived from the tumor mass.
The paper offers a wide review of the cellular techniques to test the antineoplastic drugs on tumor either in vitro cell viability tests, or organoid cultures, or patient's derived xenografts. Thees techniques are intended for every kind of tumors , noy only for STS.
On the contrary it is true that STS has few active drugs and a call to action is strictly requested
At this moment the practical consequence of functional assays on STS therapy is weak and very few data on the application of these cell drugs sensitivity are presented. No clinical study are cited or reported by the Authors and the conclusions "Personalized strategies particularly those integrating genomic and functional data provide higher potential for treatment precision, reduced toxicity, and the identification of novel therapeutic opportunities "are , in my opinion too optimistic.
In my opinion it would be useful to complete the article with a table reporting the clinical studies or trials concluded and published that confirm the value of those technique in the treatment of metastatic STS or in alternative to better define that cell functional assays are at present in the hope of clinicians involved in the treatment of STS but they aren’t of current use even in referral Center for this kind of diseases.
Nowadays , even with modest results histology -driven approach is the most accepted in sarcoma community. NGS can be an alternative solution. Functional assay is the possible future,
Author Response
Reviewer 1
Comment 1:
The article provides a good overview of functional assays in cancer but is too general and not focused enough on soft-tissue sarcomas (STS). The conclusions are somewhat optimistic given the limited clinical data.
Response:
We thank the reviewer for this valuable feedback. The revised manuscript now places greater emphasis on STS-specific data, particularly in Sections 3.1–3.3, where examples from leiomyosarcoma, liposarcoma, undifferentiated pleomorphic sarcoma, and synovial sarcoma have been integrated. The Conclusion was rewritten to reflect a more balanced interpretation, clarifying that functional assays are promising but not yet part of routine clinical practice. We also now highlight that most evidence derives from early-phase or preclinical studies, underscoring the need for prospective trials.
Comment 2:
Few or no clinical studies were cited to show how functional assays impact treatment decisions in STS.
Response:
We have incorporated references to recent translational and early clinical studies evaluating patient-derived xenografts (PDX) and in vitro cell assays that correlate ex vivo drug sensitivity with clinical outcomes in STS patients (Section 3.3). A multicenter PDX-guided treatment study is now summarized to demonstrate feasibility and translational potential, while explicitly noting that such approaches remain limited to research settings.
Comment 3:
The conclusions seem too optimistic. There are few practical consequences of functional assays for current STS therapy.
Response:
We agree and have toned down the language in the Conclusion to reflect that functional assays currently serve as research tools rather than standard diagnostics. We now explicitly state:
“Functional assays are at present a research instrument under active development. Their integration into clinical decision-making for STS will require prospective validation, workflow optimization, and standardization.”
This change ensures our conclusions are appropriately cautious and evidence-based.
Comment 4:
A table summarizing clinical studies or trials that validate these assays in STS would strengthen the paper.
Response:
While the number of clinical trials applying functional assays specifically to STS is still limited, we added a concise summary of translational and pilot clinical efforts within the text of Section 3.3, describing multicenter experiences with PDX-guided therapy. At this stage, there are no large-scale trials directly validating these assays for STS, which we clearly acknowledge in the Discussion and Conclusion as a limitation.
Comment 5:
Functional assays should be contrasted more explicitly with the currently accepted histology-driven and genomic-guided approaches.
Response:
We have revised Section 5 (Conventional Versus Personalized Therapy) to provide a detailed comparison between histology-based, genomic-based, and functional strategies. This section now discusses how hybrid algorithms combining these approaches may offer the most realistic path toward implementation in STS care.
Reviewer 2 Report
Comments and Suggestions for AuthorsThe authors generated an informative review manuscript related to a general discussion of functional assay laboratory methods utilized to guide personalized oncological treatment of patients with soft tissue sarcomas. The tables and figures are informative and support the review discussion. As the authors clearly state, the described functional assay methods are currently utilized in the research setting but are not yet ready or used in the clinical laboratory setting, although this could change in the clinical reference laboratory setting in the future. This review will likely be of interest to physicians and scientists who diagnose, manage, treat, and study soft tissue sarcomas, cancer biology, and targeted therapies. I have some minor comments to help improve the current review manuscript.
- I recommend the authors reference and discuss some specific detailed examples of published organoid and PDX model studies related to specific soft tissue sarcoma subtypes. This will provide some more relevance to soft tissue sarcomas beyond a general discussion of the organoid and PDX model functional assay methods.
- The authors should consider generating another table of soft-tissue-sarcoma cell lines as well to compliment the discussion on cell line functional studies.
- Also, since the authors discuss the tissue microenvironment, they should consider discussing studies related to digital spatial profiling in the functional analysis of organoids and PDX models of soft tissue sarcomas as well.
- Please carefully proofread the manuscript because there are occasional issues with grammar and punctuation (run-on sentences, sentence fragments, and subject-verb agreement) scattered throughout the manuscript text.
Please carefully proofread the manuscript because there are occasional issues with grammar and punctuation (run-on sentences, sentence fragments, and subject-verb agreement) scattered throughout the manuscript text
Author Response
Comment 1:
Please include more detailed examples of published organoid and PDX studies related to specific soft-tissue sarcoma subtypes.
Response:
We appreciate this helpful suggestion. Sections 3.2 (Organoids) and 3.3 (Xenografts) now include subtype-specific examples such as rhabdomyosarcoma, leiomyosarcoma, myxoid liposarcoma, synovial sarcoma, and undifferentiated pleomorphic sarcoma. Each example cites published work demonstrating how patient-derived models reproduce tumor histology, genomic fusions (e.g., SS18–SSX, FUS–DDIT3), and drug-response profiles.
Comment 2:
Add a table summarizing soft-tissue sarcoma cell lines used in functional assays.
Response:
We have addressed this by adding Table 2: “Commonly used soft tissue sarcoma (STS) cell lines in functional drug sensitivity studies” to Section 3.1.
This table summarizes key features and known sensitivities for major STS lines such as HT-1080, SYO-1, SK-LMS-1, STS26T, and others, complementing the descriptive text.
Comment 3:
Since the paper discusses the tumor microenvironment, consider mentioning digital spatial profiling (DSP) in relation to organoid and PDX models.
Response:
We fully agree. A new dedicated subsection titled “Role of Tumor Microenvironment and Digital Spatial Profiling in Organoid and PDX Models” has been added following Section 3.3.
This section explains how DSP enables spatial mapping of drug-response heterogeneity within organoids and xenografts and how its integration could enhance functional precision oncology for STS. We also note that no sarcoma-specific DSP studies have yet been published, but highlight this as a future direction.
Comment 4:
Please ensure grammatical consistency and correct minor stylistic issues.
Response:
The manuscript underwent a comprehensive proofreading and language revision to address grammar, punctuation, and style issues. All abbreviations are now defined at first mention, and figure/table numbering was standardized.
Comment 5:
Some sentences are long and could be simplified for clarity.
Response:
We have streamlined overly long sentences throughout the text and improved transitions between sections to enhance readability and flow. This edit did not alter scientific meaning but improved accessibility for both clinical and scientific readers.
