Elucidating enhancers governing human pancreatic cell identity and function

Authors

• L Wang
• J Wildenthal
• T Truongvo
• M Erdos
• H Efsun Arda

Abstract

The genetic and epigenetic regulatory logic underlying human disorders has recently been a focus for etiological models of human diseases, including diabetes mellitus. All forms of diabetes result from loss or impairment of beta-cells, main source of the principle glucose homeostasis hormone, insulin. Delineating the molecular pathways underlying the acquisition or maintenance of pancreatic cell fates and physiological functions will be instrumental to developing targeted therapies. Previously, we have used ATAC-seq and cell sorting to identify cell-type specific accessible genomic regions of normal human pancreatic cells, and revealed putative regulatory regions corresponding to specific pancreas cell lineages. However, the mechanism linking these accessible regions to gene regulation in each cell type, particularly in beta cells, remains unclear. Here, we performed Hi-C linked with chromatin immunoprecipitation (HiChIP) experiments using the EndoC cells, an insulin-producing human cell line, to elucidate the enhancer-interacting genomic regions. These data were integrated with the ATAC-seq results to determine putative enhancers unique to beta-cell function. In parallel, we established a CRISPR-interference system in the EndoC cells, featuring an efficient guide RNA delivery technique, CARGO, in order to verify and determine the functionality of these putative enhancers.

Scientific Focus Area: Genetics and Genomics

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