The roles of NRF2 in arsenic induced malignant transformation

This pilot project aims to determine the interplay between arsenic‐induced redistribution of DNA methylation sites and NRF2 activation, and how these interactions promote the associated cellular transformation. The completion of this project will not only provide a better understanding of the roles of NRF2 in arsenic‐ induced cellular transformation, it will also provide insights into how NRF2 induction by chemopreventive compounds differs from that by carcinogens. The project exploits the latest in genomic techniques to perform high‐resolution characterization of NRF2 binding sites. The requested funds will be used for the following experiments to generate preliminary data for the proposed project: 1. Model arsenic-induced cellular transformation in vitro and run a parallel control experiment using sulforaphane in place of arsenic. 2. Perform chromatin‐immunoprecipitation‐exonuclease‐sequencing (ChIP‐ Exo‐Seq) and bisulfite sequencing on the arsenic transformed and the passaged matched sulforaphane treated cells, to identify NRF2 targets that are specific to each of those NRF2 inducers. 3.  Perform molecular validations (qPCR, bisulfite‐PCR‐sanger sequencing, and luciferase reporter assay) on the identified sulforaphane specific and arsenic specific NRF2 targets.  An R01 (ES 024846-01; “The interplay between NRF2 activation and DNA methylation change in arsenic-induced carcinogenesis”) was reviewed and additional preliminary data are required to support a resubmission.