scRNA seq of in vitro co- culture of distinct subtypes of oral cancer associated primary fibroblasts and oral cancer cell line

Heterogeneity in oral cancer associated fibroblasts (CAFs)in tumor microenvironment modulate plasticity of malignant epithelial cells to drive tumorigenesis, ultimately leading to poorer clinical outcome. We identified a receptor tyrosin kinase 'TEK' or Tie2 involved in myofibroblastic differentiation of CAFs. Differentiated CAFs induce stochastic plasticity in oral cancer cells both in stemness and EMT axes. Inhibiting Tie2 in myofibroblastically differentiated CAFs purturbed their cancer cell remodelling ability. Here in this study we performed scRNA seq analysis to understand distinct CAF driven transcriptionl states of cancer cells to undertsand CAF driven cancer cells reprogramming in a higher resolution. Overall design: Oral cancer tumor derived Primary fibroblasts were cultured. Undifferentiated CAFs (C1 CAFs) were treated with TGFß for 48 hours to induce myofibroblastic differentiation. After myofibroblastic differentiation the CAFs were treated with Tie2 inhibitor. Following that, oral cancer cells were co-cultured for 96 hours with 3 different CAF phenotypes; nonmyofibroblasts (C1 CAFs), myofibroblasts (C2 CAFs), Tie2 inhibited myofibroblasts (Tie2i C2 CAFs) respectively and later harvested and scRNA-seq was performed using 10x Genomics Chromium platform