ArticleDiscover oncology2026
Integrative single-cell and spatial transcriptomic analysis reveals that ATG13 enriched in cancer-associated fibroblasts is associated with the suppression of ferroptosis in colorectal cancer.
Article in Discover oncology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
backgroundColorectal cancer (CRC) progression and metastasis are primarily driven by dynamic changes within the tumor microenvironment (TME). Cancer-associated fibroblasts (CAFs) exhibit significant heterogeneity within the TME and play a crucial role in regulating ferroptosis. Therefore, identifying novel ferroptosis-related biomarkers associated with CAFs could contribute to the development of personalized therapeutic strategies for cancer.
methodsThis study integrates single-cell sequencing data from 64 CRC samples and uses hierarchical clustering to identify cancer-associated fibroblast (CAF) subpopulations. We then combined spatial transcriptomics, hdWGCNA, CellChat, and Monocle analyses to characterize CAF subgroups. Single-cell and spatial transcriptomic data were integrated with Seurat to build a comprehensive spatial atlas. Multiplex immunohistochemical staining was used to analyze the spatial relationships of key genes with CAFs. Functional validation of these genes was performed with molecular docking, qPCR, Western blot, and both cellular and animal experiments.
resultsSingle-cell data analysis identified five distinct CAF subpopulations, among which inflammatory cancer-associated fibroblasts (iCAFs) were significantly associated with the ferroptosis pathway and closely correlated with the proliferation and metastasis of malignant epithelial cells (p = 0.87). WGCNA revealed that the iCAFs-M10 module exhibited the strongest correlation with the expression of ferroptosis-related genes (cor = 0.6796). Further investigation identified ATG13 as a key hub gene, highly expressed in tumor tissues and significantly associated with poor patient prognosis. Spatial transcriptomics and multiplex immunohistochemistry (mIHC) results demonstrated that ATG13 was highly expressed in CAFs and co-localized with CAFs markers, such as α-SMA and Vimentin. In vitro and in vivo functional assays confirmed that ATG13 knockdown effectively inhibited the proliferation, migration, and invasion of colorectal cancer cells. Additionally, ferroptosis was induced upon ATG13 silencing, as evidenced by increased levels of GPX4, SLC7A11, ACSL4, MDA, Fe
conclusionThis study identifies the iCAFs subpopulation as playing a key role in regulating ferroptosis in CRC, with ATG13 acting as a negative regulator of ferroptosis that promotes tumor progression. These findings provide new insights and potential targets for understanding the mechanisms of ferroptosis in CRC and for the development of targeted therapies.
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