Functional Characterization of Glioma GWAS Variants

胶质瘤 GWAS 变异体的功能表征

• 批准号: 9743742
• 负责人: GRAHAM CASEY
• 金额: $ 58.42万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-23 至 2022-01-25
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9743742
• 关键词: 11q23; 20q13; 3q26; 5p15.33; 8q24; 9p21; 9q21; Alleles; Autopsy; Biological; Biological Assay; Biology; Brain; Brain region; CDKN2A gene; Candidate Disease Gene; ChIP-seq; Chromatin; Chromatin Conformation Capture and Sequencing; Data; Data Set; Development; Distal; Enhancers; Epidermal Growth Factor Receptor; Event; Exons; Fluorescent in Situ Hybridization; Freezing; Future; Gene Expression; Gene Expression Profiling; Gene Targeting; Genes; Genome; Genotype; Genotype-Tissue Expression Project; Glioma; Gliomagenesis; Goals; Guidelines; Haplotypes; Hypersensitivity; Incidence; Inherited; Intervention; Knock-out; Linkage Disequilibrium; Luciferases; Malignant Neoplasms; Maps; Meta-Analysis; Methods; Molecular; Nucleic Acid Regulatory Sequences; Pathologic; Preventive; Quantitative Trait Loci; Regulatory Element; Research; Risk; Role; Sampling; Series; Single Nucleotide Polymorphism Map; Site-Directed Mutagenesis; Source; Susceptibility Gene; TNFRSF6B gene; Technology; Therapeutic; Therapeutic Intervention; Translating; Universities; Validation; Variant; base; brain tissue; causal variant; chromosome conformation capture; experience; genome editing; genome wide association study; genome-wide; glioma cell line; improved; insight; novel; promoter; risk variant; success; tool; transcriptome sequencing; vector

PROJECT SUMMARY/ABSTRACT The goal of the proposed study is to discern the functional and biological relevance of gliomas risk variants identified through genome wide association studies (GWAS). GWAS have led to the discovery of 8 susceptibility loci in glioma: 8q24.11, 11q23.3, 5p15.33, 9p21.3, 20q13.33, 7p11.2 (2 independent loci) and 3q26.2. None of the GWAS SNPs map to exons and we hypothesize that the associated SNPs are non- functional, are in linkage disequilibrium with casual/functional SNPs, and map to risk enhancers that in turn regulate target gene expression in an allele specific manner. To date no functional/causal variants for glioma GWAS have been identified. However, our team has identified some candidate target genes for 6 of the 8 GWAS loci using expression quantitative trait loci (eQTL) mapping in multiple brain regions and allelic specific gene expression (ASE) analyses. Based on these promising findings, we propose a comprehensive post GWAS analysis of glioma risk loci using a series of complementary approaches. In Aim 1 we will identify candidate target genes of glioma risk loci using data from two sources: publicly available RNA-Seq and genotyping data from multiple brain regions of 400 post-mortem brains of the Genotype-Tissue Expression Project's (GTEx); we will also generate RNA-Seq and genotyping data from an additional 300 pathologically verified, fresh-frozen autopsied normal brain tissues (multiple brain regions) from the University of Miami Brain Bank (UMBB). eQTL mapping, eQTL meta-analyses and eQTL-ASE will be performed, using the UMBB samples as the discovery set and GTEx dataset as a validation dataset. In Aim 2 we will use publicly available ChIP-Seq and DNAse1 hypersensitivity chromatin data to identify candidate regulatory elements within GWAS loci. We will: (a) validate candidate regulatory elements using enhancer/promoter luciferase vector activity assays in multiple glioma cell lines. (b) Assess allele specific effects on enhancer/promoter activity using either site-directed mutagenesis or naturally occurring haplotypes (c). Identify and validate novel candidate target genes of risk enhancers identified in Aim 1 by knocking out risk enhancers using CRISPR-Cas9 gene editing technology in glioma cell lines and assessing changes in target gene expression using RNA-Seq. In Aim 3 we will use existing data and novel data generated through Aims 1 and 2, to identify and validate the physical interaction between risk enhancers and candidate target genes. Interacting loci of candidate risk enhancers will be identified using 4C-Seq using five glioma cell lines, and will be compared to candidate target genes identified through Aims 1 and Aim 2. Any significant interactions, especially those with candidate local and distal target genes identified through Aims 1 and 2 will be further validated by 3C and fluorescent in situ hybridization (FISH). Through these efforts we will develop a mechanistic and biological understanding of glioma risk that will be an essential first step in our translational efforts to develop preventive therapeutics approaches for glioma.

项目总结/文摘

项目成果

A Functional Variant on 9p21.3 Related to Glioma Risk Affects Enhancer Activity and Modulates Expression of CDKN2B-AS1.

• DOI: 10.1002/humu.24244
• 发表时间: 2021-10
• 期刊: Human mutation
• 影响因子: 3.9
• 作者: Ali MW;Patro CPK;Devall M;Dampier CH;Plummer SJ;Kuscu C;Adli M;Lai RK;Casey G
• 通讯作者: Casey G