Profiling of epithelial and stromal cells in early proximal colon preneoplasia with matched normals.

Progression of nonmalignant colonic cells harboring somatic mutations depends upon interactions with surrounding stroma; however, details of this interaction are poorly understood. We performed targeted transcriptional profiling of laser-captured epithelial and stromal cells from ACF, the earliest detectable human colonic pre-neoplastic lesion. Within ACF epithelium carrying non-overlapping mutations in KRAS, BRAF, or APC, we observed broad NF-kB-related inflammatory changes and specific differences associated with each mutation, including BRAFV600E-mediated senescence. Additionally, we identified ACF-associated stromal changes indicative of immune cell infiltration and fibroblast activation. Our results provide new insight into intercellular signaling changes that control the earliest stages of CRC development. Epithelial and stromal targetted mRNA profiles of biopsied proximal colon ACF and normal samples. Each ACF had a detected mutation for either KRAS, BRAF, or APC via DNA-MS. Epithelial and stromal cells were individually isolated using laser capture microdissection. mRNA profiles were evaluated using two Ampliseq RNA panels (off the shelf Apoptotic panel, and custom Senescence panel of 20 genes)