The Epitranscriptome as a Novel Mechanism of Arsenic-Induced Diabetes.

PROJECT SUMMARY The parent grant, entitled “The Epitranscriptome as a Novel Mechanism of Arsenic-Induced Diabetes”, aims to investigate the impacts of low-level arsenic (As) exposure on the N6-methyladenosine (m6A)-epitranscriptome and on diabetes in American Indian communities in the US. The parent study proposed to test the hypothesis that altered m6A is plausible mechanism for As-related diabetes in the Strong Heart Study (SHS) cohort. Our original proposal relied on immunoprecipitation-based sequencing methods that required large amounts of RNA and were unable to profile m6A at the single base resolution. However, recent technological advances by Oxford Nanopore Technologies (ONT) now allow for direct detection of m6A on RNA strands, improving RNA input and providing invaluable information on transcript splicing and isoforms, along with other modifications to RNA. We are requesting a supplemental award to enable the best possible science for our Aims, given the rapid and significant changes in the chemistry, protocols, and informatics capacity of the direct RNA sequencing methods. Specifically, in our hands, the latest flowcells and chemistry enable a dramatic increase in accuracy (from 75% to 98%), new tools for calling base modifications (i.e. m6A) and better throughput. However, the new flowcells and chemistry are more expensive, and would necessitate additional funding beyond the parent award to enable processing of all the banked samples from our study with the best methods for RNA modification detection and epitranscriptome mapping of environmental exposure.