Review for 10.1186/s12876-025-03920-0 (2025)

Title: Unveiling the heterogeneity and immunotherapy potency of tumor-associated neutrophils
in the tumor microenvironment of Gastric cancer
Version: 1 Date: 09 Mar 2025
Reviewer:  Anwar Ashraf Abouelnasr
Reviewer's report:
Dear Authors, please address the following points below
-While the manuscript does mention clinical data analysis (e.g., ROC analysis and survival data),
there could be a more detailed interpretation of how the findings from the scRNA-seq analysis
translate into clinical outcomes. For example, how do the identified TAN subpopulations
correlate with patient survival, metastasis, or response to immunotherapy? The study could
benefit from deeper insights into how TANs affect clinical variables.
-The manuscript focuses heavily on neutrophils but provides less context regarding the broader
immune cell landscape within the TME. A more comprehensive discussion of how TANs
interact with other immune cells (e.g., T cells, macrophages) and their collective role in shaping
the immune response in GC would provide a more holistic understanding of the TME and its
influence on immunotherapy outcomes.
-The study primarily relies on the GEO datasets, which may have inherent biases, such as
patient selection or tissue sample heterogeneity. While multiple data sources are used, the
generalizability of the results could be better addressed. For example, the study could incorporate
validation from independent external cohorts or include more clinical data to strengthen the
relevance of findings in real-world GC patients.
-The analysis heavily relies on computational methods and bioinformatics tools to predict the
role of TANs in GC. While these methods are powerful, experimental validation (e.g., flow
cytometry, immunohistochemistry, or in vivo models) would strengthen the claims made in the
manuscript and provide experimental evidence supporting the findings. In particular, validating
the identified TAN subpopulations and their therapeutic potential in animal models would add
substantial weight to the findings.
-While the study identifies novel neutrophil-related targets for immunotherapy, the manuscript
does not sufficiently discuss other emerging immune targets for GC. Given the rapid
advancement of immune therapies targeting different immune cells (e.g., macrophages, T
cells), it would be valuable to contextualize the role of TANs within the larger landscape of
immunotherapy research in GC.
-Although the manuscript identifies various subtypes of TANs and their potential interactions
within the TME, there is minimal functional characterization of how these neutrophils impact
tumor progression, metastasis, or immune suppression in GC. Experimental work on TAN
functions (e.g., their ability to form neutrophil extracellular traps or their role in metastasis)
would provide a clearer understanding of their role in GC pathophysiology.
-While the paper proposes the role of specific TAN subtypes in GC progression, there is a lack
of robust in vivo and in vitro validation to support these claims. Experimental data from both
animal models and human tissue samples would significantly strengthen the conclusions drawn
about TANs’ involvement in tumor progression and metastasis.
-While the paper identifies key pathways and genes involved in TAN behavior, the mechanistic
details of how these pathways contribute to tumor progression are still somewhat vague. For
example, while RGS2 is highlighted as a potential target, further exploration of its functional
role in GC would be helpful to justify its therapeutic potential fully.
-While the study emphasizes the role of TANs, it could also integrate a broader view of the TME
by considering interactions between TANs and other immune cell types. Understanding how
neutrophils cooperate or antagonize other immune cells, such as T cells, macrophages, and NK
cells, would offer a more holistic view of the immune landscape in GC.