Bulk_RNA_HepG2-sc-6250
Bulk_RNA_HepG2
Recent work in the Campbell group had uncovered somatic mutations present in normal, non-cancerous human liver tissue. Detectable driver mutations likely confers a selective advantage to the affected cells, leading to clonal expansions, likely raising the risk of progression of chronic liver disease, potentially contributing to the onset and recurrence of hepatocellular carcinoma. Understanding the downstream consequences of such somatic driver mutations in liver tissue is important to advance our understanding of diseases/malignancies affecting the liver including diabetes and tumorigenesis and could inform the design of follow-on studies leading to novel mechanistic insights that could be clinically relevant.   In this project, our goal is to evaluate the differences in RNA abundance by RNA-Sequencing among cells with versus without driver somatic mutations in the FOXO1 transcription factor in the HepG2 cell line, across a range of nutrient-controlled conditions (i.e., starved vs insulin-stimulated). Subsequent analysis of differences in biological pathway activity and gene expression programs will be quantitatively assessed across the experimental conditions considered.
Chronic liver disease is associated with metabolic dysregulation, liver failure and hepatocellular carcinoma. We analyzed somatic mutations from 1202 genomes across 32 liver samples, including normal controls, alcohol-related and non-alcoholic fatty liver disease. Five of 27 patients with liver disease carried hotspot driver mutations in FOXO1, the major transcription factor downstream of insulin signaling. FOXO1 mutations were independently acquired by up to 5 distinct clones within the same patientâ€™s sample, and impaired insulin-mediated nuclear export of FOXO1. GPAM, which produces storage triacylglycerol from dietary calories, also had significant excess of mutations, similarly exhibiting convergent evolution within biopsies. Telomeres were shorter in diseased than normal liver, with attrition more pronounced in larger clones. Multiple independent acquisitions of drivers within one small liver sample imply that such mutations could affect hundreds of grams of tissue across the whole organ, potentially contributing to systemic metabolic dysfunction. The downstream consequences of the observed driver mutations will be assessed using RNA-Seq experiments in the HepG2 cell line.