Abstract PR012: Aberrant cell state plasticity mediates colorectal cancer initiation

Abstract

Cell state plasticity – the ability of cells to acquire new states via differentiation programs - is an important feature of embryogenesis, allowing for tissue specification during development, and adult homeostasis, enabling adaptive responses to physiological cues and pathological states. Epigenetic governance and gene regulation endow epithelia with restricted cell state plasticity that enable tissue homeostasis while avoiding properties that may facilitate neoplasia. There is emerging evidence that aberrant expansion of the normally restricted capability for cell state plasticity to escape terminal differentiation is a key aspect of cancer initiation. The nongenetic factors and specific programs that mediate aberrant cell state plasticity require deeper characterization to understand this elusive aspect of cancer pathogenesis. Previously challenging to study, cell state plasticity can now be investigated using single cell technology, offering unprecedented molecular resolution. We applied state-of-the-art methods on primary human tissue and derivative organoids as well as different mouse models to (a) characterize aberrant transcriptional cell states enabling cancer initiation and (b) define a functional mediator of expanded cell state plasticity. Using genetically engineered and carcinogen-induced murine models of intestinal neoplasia, we demonstrate that inappropriate Sox9 expression and loss of post-mitotic villus differentiation are early events preceding cancer development. By applying single cell RNA-sequencing (scRNA-seq) to flow-sorted epithelial cells from the Lgr5Cre; Apcf/f; R26tdT genetic model, we observed that preneoplastic cells express specific intestinal stem cell genes but are otherwise transcriptionally rewired, evading resemblance to normal intestinal cell types; we therefore refer to these cells as aberrant stem cell-like (AbSC). We further analyzed the AbSC transcriptional state, searching for features that explain its distinguished gene expression profile, and found that it is characterized by impaired differentiation, enhanced regenerative capacity, and reactivation of fetal genes. The evaluation of preneoplastic colonic lesions from the carcinogen-induced mouse model and a patient with familial adenomatous polyposis (FAP) by scRNA-seq confirmed these findings. Notably, while chromatin accessibility increased at regenerative genes, new accessibility was observed at fetal intestinal genes by ATAC-seq. Genetic inactivation of Sox9 prevented adenoma formation in ApcKO mice, obstructed emergence of the aberrant transcriptional state, including genes reserved for fetal intestinal development, and restored multi-lineage differentiation by scRNA-seq. SOX9 knockdown in human adenoma organoids compromised expression of fetal genes and induced differentiation. These studies indicate that cancer initiation requires aberrant phenotypic plasticity mediated by unabated regenerative activity and developmental reprogramming. Citation Format: Pratyusha Bala, Jonathan Rennhack, Clare Morris, Daulet Aitymbayev, Sydney M. Moyer, Gina N. Duronio, William Hahn, Nilay S. Sethi. Aberrant cell state plasticity mediates 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 PR012.