Molecular Pathogenesis of early colorectal cancer through investigation of the mechanism of colitis to cancer progression

Inflammatory bowel disease is a significant risk factor for the development and progression of colorectal cancer.  Chronic inflammation orchestrates intestinal epithelia transformation that culminates in malignancy. Hyperactivated immune cells, inflammatory cytokines, and oxidant stress have been shown to result in mutations and progression to malignancy. The aim of this proposal is to determine the molecular profiles of that infer progression from IBD to CRC. The initial focus is to study the role of the CBM signalosome complex which is key part of the NFkB pathway and consists of three key molecules; BCL10, MALT1, and CARD11. Human FFPE tissue biopsies from IBD and CRC patients will be subjected to immunohistochemistry to characterize BCL10, CARD11 and MALT1 their protein expression. In addition, functional validation of the CBM complex will be carried out using HT29 and HCT116 cell lines.  This was carried out by cloning the CARD11 gene in pcDNA3.1 expression vector, followed by  assessment of the expression of CARD11 using western blotting and qRT-PCR. Preliminary result from cultured cells indicates that transfection resulted in increased levels of CARD11 expression in transfected cells when compared to the control. Characterization of the role of CARD11 in early CRC pathogenesis will be carried out using whole transcriptomic analysis of  FFPE patient samples and the transfected CRC cell lines compared CR cell lines. This will be followed by studying the role of BCL10 and MALT1 in similar manner. These molecules are implicated in CRC progression.