Gene expression profiles of interactions between colorectal cancer-associated fibroblasts and myeloid cells in 2D co-cultures and in novel next generation triple cultures containing colorectal cancer patient-derived organoids

In this study colorectal cancer (CRC) patient-derived cancer-associated fibroblasts (CAFs) and primary monocytes or macrophages were co-cultured in 2D set-up and single-cell transcriptomics was employed to analyze the phenotypic and functional changes of the myeloid cells and CAFs upon their interaction. The results showed that CAFs instruct monocytes to gain a tumor-associated macropages (TAM)-like cells phenotype. In the next step, novel next generation tumor organoid cultures of myelid cells, CAFs and patient-derived CRC organoids (PDOs) were utilized to mimic the tumor microenvironment and study the phenotypic changes of cells under chemotherapy and oncolytic virus treatments. The sequencing data revealed that both treatments triggered distinct phenotypic changes and identified immunomodulatory effects of chemotherapy in TAM-like cells. Oncolytic virus treatment led to strong upregulation of Type I and II interferon signaling, while both therapies activated multiple cell-cell communication pathways, highlighting key molecular interactions relevant to the tumor microenvironment. This co-culture model provided insights into TAM-like responses, aiding precision cancer treatment evaluations.