Impact of disease associated noncoding polymorphisms on post-transcriptional gene regulation

疾病相关非编码多态性对转录后基因调控的影响

• 批准号: 9246421
• 负责人: Srinivas Mummidi
• 金额: $ 36.51万
• 依托单位: UNIVERSITY OF TEXAS RIO GRANDE VALLEY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9246421
• 关键词: 3' Untranslated Regions; Affect; Alleles; Antigens; Atherosclerosis; Attention; Autoimmune Process; Binding; Biological Models; CRISPR/Cas technology; Cells; Clinical; Communicable Diseases; Crohn' s disease; Development; Disease; Disease Outcome; Disease susceptibility; Electrophoretic Mobility Shift Assay; Elements; Enhancers; Exhibits; Fractionation; Gene Expression; Gene Expression Regulation; Genes; Genetic Polymorphism; Genetic Transcription; Genetic Variation; HIV; HU Protein; Haplotypes; HuR protein; Human; Immune; Immunoprecipitation; In Vitro; Inflammatory; Knowledge; Ligands; Link; Linkage Disequilibrium; Maps; Mediating; Mediator of activation protein; Messenger RNA; Molecular and Cellular Biology; Nucleic Acid Regulatory Sequences; Pathogenesis; Pathology; Play; Polyribosomes; Predisposition; Probability; Proteins; Publications; RNA; RNA Binding; RNA Interference; RNA-Binding Proteins; Records; Recruitment Activity; Risk; Risk stratification; Single Nucleotide Polymorphism; Site; Structure; T-Lymphocyte; Technology; Testing; Therapeutic Agents; Transcript; Untranslated Regions; Variant; base; beta-Chemokines; genetic variant; genome wide association study; induced pluripotent stem cell; innovation; mRNA Expression; mRNA Stability; macrophage; monocyte; new therapeutic target; overexpression; promoter; public health relevance; stem; targeted treatment; technological innovation; trait

 DESCRIPTION (provided by applicant): Genome-wide association studies (GWASs) have identified multiple disease (trait) associations. While many variants map to 5' cis-regulatory regions (e.g. enhancers), many also localize to the 3'-untranslated region (UTR), suggesting that post-transcriptional mechanisms such as mRNA stability and translatability may play a key role in disease susceptibility. Supporting such a possibility we show that a disease-modifying haplotype in the CC chemokine ligand 2 (CCL2) 3'UTR is associated with allelic expression imbalance (AEI), increased mRNA stability and increased binding to Hu Antigen R (HuR), a RNA stabilizing protein. Notably this haplotype designated as rs1024611G-rs13900T is associated with a number of inflammatory diseases that are associated with monocyte/macrophage recruitment. Our overarching hypothesis is that functional genetic variation in untranslated regions is associated with altered mRNA stability/translatability which determines gene expression and impacts disease susceptibility and pathogenesis. Specific Aim 1 tests the hypothesis that the disease-influencing rs1024611G-containing allele augments CCL2 mRNA expression via post-transcriptional mechanisms, resulting from enhanced binding of the RNA stabilization protein Hu Antigen R (HuR) to the CCL2 3'-UTR. We will rigorously test the hypothesis that the rs13900T-bearing haplotype is linked to mRNA stability and/or altered mRNA translatability mediated via HuR by AEI analyses, RNA mobility shift assays, allele specific RNA immunoprecipitation, RNA interference, overexpression of RNA binding proteins (RBP) and polysome fractionation in primary macrophages and in isogenic macrophages differentiated from iPSC. Significance: rs1024611G-rs13900T haplotype impacts disease pathogenesis in many inflammatory diseases. Specific Aim #2 extends the concepts of aim 1 and examines the functional (AEI) and post-transcriptional impact (differential binding of RBP, mRNA stability/translatability) of SNPs in the 3'-UTR of genes that by GWAS are highly associated with disease susceptibility. These candidate polymorphisms are selected based on their strong disease associations and high probability of regulatory function. The proposed studies will greatly expand our understanding of post-transcriptional mechanisms mediated through genetic variation in inflammatory genes. Innovation, Significance and Clinical Impact: Identification of functional SNPs remains a primary challenge in post-GWAS era. Hitherto, particular emphasis has been placed on genetic variants in promoters and enhancers to ascribe function for GWAS identified genetic variants. The conceptual idea that disease-influencing SNPs affect disease outcomes through post-transcriptional mechanisms is innovative. Such studies are of high translational value as they will facilitate risk-stratification, early intervenion and development of targeted therapies.

 描述（由适用提供）：全基因组关联研究（GWASS）已确定多种疾病（特征）关联。尽管许多变体映射到5'顺式调节区域（例如增强剂），但许多变体也定位于3'-非翻译区域（UTR），这表明诸如mRNA稳定性和翻译性之类的转录后机制可能在疾病易感性中起关键作用。支持这种可能性，我们表明，CC趋化因子配体2（CCL2）3'UTR中的疾病改良单倍型与等位基因表达不平衡（AEI）相关，mRNA稳定性增加并增加了与Hu抗原R（HUR）的结合（一种RNA稳定蛋白）。值得注意的是，该单倍型被指定为RS1024611G-RS13900T与与单核细胞/巨噬细胞募集相关的许多炎症性疾病有关。我们的总体假设是，未翻译区域的功能遗传变异与mRNA稳定性/转换性的改变有关，这决定了基因表达并影响疾病的易感性和发病机理。具体目的1检验了以下假设：含有RS1024611G的RS1024611G等位基因通过转录后机制增强了CCL2 mRNA的表达，这是由于RNA稳定蛋白HU抗原R（HUR）对CCL2 3'-UTR的结合而引起的。我们将严格检验以下假设：含RS13900T的单倍型与mRNA的稳定性和/或通过AEI分析，RNA迁移率转移，等位基因RNA特定的RNA免疫沉淀，RNA免疫应对，RNA的过度表达，RNA对RNA结合蛋白的过度表达，RNA迁移分析和/或改变的mRNA转换性改变了RNA的迁移分析（RNA）。与IPSC区分开。意义：RS1024611G-RS13900T单倍型会影响许多炎症性疾病的疾病发病机理。具体目标＃2扩展了目标1的概念，并检查了在GWAS的3'-UTR中，SNP的功能（AEI）和转录后影响（RBP的差异结合，mRNA稳定性/可翻译性）与GWAS与疾病易感性高度相关的基因。这些候选多态性是根据它们的强疾病关联和调节功能的高可能性选择的。拟议的研究将大大扩展我们对通过炎症基因遗传变异介导的转录后机制的理解。创新，意义和临床影响：功能性SNP的识别仍然是GWAS后时代的主要挑战。迄今为止，特别重点已放在启动子和增强子中的遗传变异上，以分配GWAS鉴定的遗传变异的功能。概念上的想法是，通过转录后机制影响疾病的SNP会影响疾病的结果。这样的研究具有很高的转化价值，因为它们将促进靶向疗法的风险分层，早期和发展和发展。