Genetic variation influencing brain AVM

脑动静脉畸形（AVM）是年轻人颅内出血（ICH）的重要原因， 成年人了病因和发病机制是未知的，更好地了解信号通路 需要影响疾病易感性和临床进展以优化患者护理。在当前 在资助期间，我们取得了几项新颖而重要的发现，报告了有史以来第一个基因 与AVM易感性和颅内出血（ICH）相关。这些发现为工作奠定了基础 在下一阶段，我们将研究遗传变异对疾病易感性的影响， 使用AVM病例的大量现有DMA收集（n=647）的集中和全面的方式，以及 预计在项目过程中将入组新病例（n=370），总样本量为1017 例 目的1是一项全基因组关联（GWAS）研究，以确定与以下相关的新候选变体： AVM，使用AffyssSNP Array 6.0，包含约100万个SNP和约100万个拷贝数探针 变异（CNV）。主要分析将在我们最大的亚组中比较AVM病例与健康对照， 高加索人（520例和643例对照），使用新的基因组方法对病例和对照进行基因匹配。 控制和调整人口分层使用主成分分析。次要分析将 评估CNV是否与AVM易感性相关。 Aim 2将通过精确定位与易感性相关的候选变异， 20个含有来自GWAS分析的最高度显著的SNP和/或CNV的靶区域，通过 各种标准方法。我们的顶级相关候选SNP将在AVM病例和对照中进行验证 来自波恩大学（德国）和乌得勒支大学（荷兰）。其他分析将测试 AVM病例中的其他人种-种族组和至随后出血的时间。此外， 将使用标准分子技术检测AVM相关变体。 目的3将探索体细胞变异作为AVM的遗传机制，使用CNV分析， 通过检查杂合性缺失，比较基因组DNA与从手术标本中分离的损伤DNA 或等位基因失衡。在100例患者中，我们将对从AVM中分离的血管组织进行DNA基因分型 在手术中用Affyellow 6.0阵列去除病灶，并比较生殖系DNA之间的全基因组CNVs 和病变DNA，后续将包括使用亚克隆和深度重测序。

Brain arteriovenous malformations (AVMs) are an important cause of intracranial hemorrhage (ICH) in young adults. The etiology and pathogenesis are unknown and better understanding of the signaling pathways influencing disease susceptibility and clinical progression is needed to optimize patient care. In the current funding period, we have made several novel and important discoveries, reporting the first-ever genetic associations with AVM susceptibility and intracranial hemorrhage (ICH). These findings set the stage for work during the next period, during which we will study the influence of genetic variation on disease susceptibility in a focused and comprehensive fashion using a large existing DMA collection from AVM cases (n=647) and expected new cases to be enrolled (n=370) over the course of the project, yielding a total sample size of 1017 cases. Aim 1 is a Genome-wide association (GWAS) study to identify novel candidate variants associated with AVM, using the Affymetrix SNP Array 6.0, containing -1 million SNPs and ~1 million probes for copy number variation (CNV). The primary analysis will compare AVM cases to healthy controls in our largest sub-group, Caucasians (520 cases and 643 controls), using novel genomic methods to genetically match cases and controls and adjust for population stratification using principal components analysis. Secondary analysis will evaluate whether CNV is associated with AVM susceptibility. Aim 2 will follow-up candidate variants associated with susceptibility through fine-mapping of approximately 20 target regions containing the most highly significant SNPs and/or CNVs from GWAS analysis through a variety of standard methods. Our top-associated candidate SNPs will be validated in AVM cases and controls from the University of Bonn (Germany) and University of Utrecht (Netherlands). Additional analyses will test other race-ethnic groups and time-to-subsequent hemorrhage in AVM cases. Further, biological function of AVM-associated variants will be tested using standard molecular techniques. Aim 3 will be exploration of somatic variation as a genetic mechanism in AVM using CNV analysis to compare genomic DNA to lesional DNA isolated from surgical specimens by examining loss of heterozygosity or allelic imbalance. In 100 patients, we will genotype DNA from vascular tissue that is dissected from the AVM nidus removed at surgery with Affymetrix 6.0 arrays, and compare CNVs genome-wide between germline DNA and lesion DNA, and follow-up will include use of subcloning and deep re-sequencing.