The Effect of High Fat Diet on Peroxisome Proliferator Activated Receptor and NF-kappaB Enhancer Region Activity

A high fat diet (HFD) has been established as a risk factor for tumor formation in mammalian tissues. In the intestine, this tumor formation may arise due to enhanced self-renewal and proliferation of intestinal stem cells (ISC). Previous studies have demonstrated that the PPARα and PPARδ isoforms play a role in activating fatty acid oxidation metabolic programs which contribute to the observed high fat diet phenotype. Computational transcriptomics, particularly ATAC-seq, has been used to identify regions which demonstrate increased or decreased accessibility as an effect of a HFD. In this study, we target motifs from CIS-regulatory regions which demonstrated increased accessibility, specifically stimulating computationally predicted PPARδ enhancer regions. This study measures activation of these enhancer regions through transduction of dual fluorescence lentiviral constructs into small intestine-derived organoids, measuring RFP and GFP levels through fluorescence microscopy. Preliminary data does not demonstrate changes in fluorescence upon stimulation.