Identifying regulatory uORFs as a targetable axis for hereditary disease

Abstract The aim of this grant is to identify and validate for therapeutic intervention, regulatory elements within the 5’ untranslated regions (UTR) of protein coding genes, known as upstream open reading frames (uORFs). In so doing, we aim to modulate the protein output from selected genes, offering a novel, translational approach with broad potential. To achieve the goals of our integrative MultiPI R01, we will leverage the expertise of both our labs spanning computational modeling, genomics, genetics, RNA biology and molecular biology. In Aim 1 we will combine large genomic and genetic datasets including gnomAD, the PennMedicine BioBank, and the UKBioBank to systematically identify functional uORFs and prioritize those for validation. The resulting database of human uORFs will be made publicly available and widely accessible as a user-friendly web-tool. Predicted regulatory uORFs suggested by the Aim 1 pipeline will be fed into the experimental Aim 2. In Aim 2 we will validate the high-priority uORF targets using luciferase assays. The results of Aims 2 will be fed back to the pipeline of Aim1 to improve prioritization of future uORFs. Overall, we expect the resources and discoveries made by this grant to shed light on the functional role of uORFs and offer therapeutic avenues for several hereditary diseases.