Abstract B023: Sox9 drives an aberrant transcriptional program impeding intestinal differentiation in colorectal cancer initiation

Abstract

Abstract Despite the implementation of screening and preventative strategies, colorectal cancer (CRC) remains the second most deadly cancer worldwide and is responsible for an alarming trend of increasing prevalence among younger patients. Inappropriate WNT activation is the initiating step in sporadic CRC development, typically through deleterious mutations in the pathway negative regulator Adenomatous polyposis coli (APC). WNT pathway hinders whereas transforming growth factor (TGF-β)/bone morphogenetic protein (BMP) signaling supports differentiation of progenitors into mature enterocytes, establishing a crypt-villus gradient. Genomic alterations that dysregulate intestinal stem cell differentiation are central to CRC development. Impairment of differentiation and inappropriate Sox9 overexpression are conserved events in CRC initiation based on the evaluation of two genetically engineered mouse models, two carcinogen-induced mouse models, and a patient with familial adenomatous polyposis (FAP), a hereditary polyposis syndrome in which a mutant copy of APC is inherited. Single cell RNA-sequencing (scRNA-seq) of adenomas from an ApcKO mouse model and FAP sample implicates an aberrant stem cell-like program (herein referred to as AbSC) as a key aspect of CRC initiation. The AbSc program is characterized by selective intestinal stem cell activity, indiscriminate attenuation of differentiated lineages, and aberrant activation of genes associated with fetal intestinal development. Histopathology and scRNA-seq of adenomas and derivative organoids from a patient with FAP demonstrated a partial block in differentiation and confirmed reactivation of genes reserved for intestinal development. Assay for Transposase-Accessible Chromatin with high-throughput sequencing (ATAC-seq) of APCKO epithelial cells revealed increased chromatin accessibility at fetal intestinal genes. Genetic inactivation of Sox9 prevented adenoma formation in ApcKO mice and induced differentiation of ApcKO organoids, obstructing the emergence of the ApcKO transcriptional state, including reactivation of a fetal-like intestinal program, restored multi-lineage differentiation, and markedly reduced the gained chromatin accessibility at developmental genes in neoplasia. These studies indicate that an aberrant transcriptional state hindering intestinal differentiation mediated by Sox9 is an early conserved event in CRC and carry important implications for developing therapeutics directed at inducing intestinal differentiation. Citation Format: Pratyusha Bala, Jonathan P. Rennhack, Clare Morris, Daulet Aitymbayev, Sydney M. Moyer, Gina Nicole Duronio, Paul Doan, William C. Hahn, Nilay S. Sethi. Sox9 drives an aberrant transcriptional program impeding intestinal differentiation in colorectal cancer initiation [abstract]. In: Proceedings of the AACR Special Conference on Colorectal Cancer; 2022 Oct 1-4; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_1):Abstract nr B023.