Identifying regulatory uORFs as a targetable axis for hereditary disease

摘要 这笔赠款的目的是确定、验证、然后针对治疗干预、 蛋白质编码基因的5‘非翻译区，称为上游开放阅读框架(UORF)。 在这样做的过程中，我们的目标是调节选定基因的蛋白质输出，提供一种新颖的、翻译的 具有广泛潜力的方法，我们将首先在遗传病动物模型的背景下进行测试。至 为了实现我们的集成MultiPI R01的目标，我们将利用我们两个实验室的专业知识 计算建模、基因组学、遗传学、RNA生物学、分子生物学和动物工作。在目标1中，我们 将结合大型基因组和遗传数据集，包括gnomAD、PennMedicine Biobank和 UKBioBank系统地识别功能性uORF，并对那些进行验证的优先顺序。由此产生的 人类uORF数据库将作为一个方便用户使用的网络工具公开提供和广泛使用。 由目标1管道建议的预测的调控uORF将被输入实验目标2和3。 目标2我们将使用荧光素酶分析来验证高优先级uORF靶标，在目标3中我们将测试 这些uORF的调节作为治疗干预的潜力。AIMS 2和AIMS 3的结果将被提供 回到Aim1的管道中，以改进未来uORF的优先顺序。总体而言，我们预计资源和 这笔赠款的发现阐明了uORF的功能作用，并为 几种遗传性疾病。

Abstract The aim of this grant is to identify, validate, then target 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, that we will first test in the context of animal models of hereditary diseases. 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, molecular biology, and animal work. 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 Aims 2 and 3. In Aim 2 we will validate the high-priority uORF targets using luciferase assays and in Aim 3 we will test the modulation of these uORFs as a potential for therapeutic intervention. The results of Aims 2 and 3 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 uORF and offer therapeutic avenues for several hereditary diseases.