IDENTIFICATION OF EPIGENETIC VULNERABILITIES IN THE ADENOMA-CARCINOMA SEQUENCE

Abstract

Abstract Introduction Numerous studies have documented the functional effect of different driver mutations on the development and progression of colorectal cancer (CRC). However, the molecular-level impact of each of these mutations on the epigenome/transcriptome, as well as the phenotype-related longitudinal effects of these mutations have not been characterized in detail. This work proposes an experimental design based on the integration of different -omic layers in order to identify molecular vulnerabilities with therapeutic potential in the adenoma-carcinoma sequence. Methods The proposed experimental model contemplates the introduction of sequential mutations in the APC/KRAS/TP53 and SMAD4 genes with CRISPR/Cas9 technology in human colon organoids obtained from healthy mucosa. To determine the molecular alterations at each step of the sequence, a paired RNA expression and DNA methylation analysis will be performed at the whole genome level with the respective massive next-generation sequencing technologies / EPIC arrays. Finally, to identify possible functional associations between the epigenome/transcriptome, an integration analysis of these -omics will be performed with novel computational algorithms, also including the integration of these data with information available from international consortia. Results The project is currently in the initial stages of development. The model and the most updated results available to date will be discussed in detail during the congress. Conclusions These results will allow the identification of molecular vulnerabilities with therapeutic potential in the context of CRC.