Project 3: Contribution of Non-VWF Genomic Variants to Quantitative Von Willebrand Factor Pathologies

项目 3：非 VWF 基因组变异对定量冯维勒布兰德因子病理学的贡献

• 批准号: 10379437
• 负责人: DAVID P LILLICRAP
• 金额: $ 23.46万
• 依托单位: VERSITI WISCONSIN, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-15 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10379437
• 关键词: Anabolism; Area; Attention; Binding; Binding Proteins; Biology; Blood; Blood Coagulation Disorders; Blood Platelets; C-Type Lectins; C2 Domain; CRISPR/Cas technology; Cells; Chromatin Structure; Code; Collagen; Consensus; Deletion Mutation; Development; Disease; Distant; Elements; Endocytosis; Endothelial Cells; Factor VIII; Family; Genes; Genetic; Genetic Enhancer Element; Genetic Transcription; Genetic Variation; Genetic study; Genome; Genomic approach; Genomics; Genotype; Hemorrhage; Human; Inherited; Knock-out; Knockout Mice; Knowledge; Ligand Binding; Mediating; Megakaryocytes; Modification; Mus; Nuclear; Nucleic Acid Regulatory Sequences; Pathogenicity; Pathologic; Pathology; Pathway interactions; Patients; Phenotype; Plasma; Polysaccharides; Population; Prevalence; Process; Proteins; RNA Splicing; Regulation; Role; Signal Pathway; Single Nucleotide Polymorphism; Site; Symptoms; System; Testing; Transcriptional Activation; Transgenic Mice; Variant; Vascular Endothelial Cell; base; cell type; cis acting element; cohort; disease phenotype; experimental study; genetic variant; genome wide association study; genomic locus; humanized mouse; in vitro Assay; in vitro Model; in vivo; indexing; member; molecular pathology; mouse model; patient population; programs; promoter; rare condition; rare variant; receptor; receptor expression; receptor mediated endocytosis; study population; syntaxin; syntaxin-2; trafficking; trait; von Willebrand Disease; von Willebrand Factor

PROJECT SUMMARY: PROJECT 3 The pathogenic basis of the inherited bleeding disorder, von Willebrand disease (VWD), represents a spectrum of genetic mechanisms and variants. While the qualitative type 2 forms of VWD are the result of missense substitutions in domains of von Willebrand factor (VWF) involved in ligand binding, the quantitative traits, type 1 and 3 VWD, are associated with many different sequence variants throughout the VWF gene. However, following detailed genotyping of the VWF coding region and splices sites in >500 type 1 and >100 type 3 VWD cases, pathogenic variants have not been identified in ~35% and ~15% of patients, respectively. This evidence, along with repeated results from large genome wide association studies (GWAS), strongly supports the involvement of genes in addition to VWF in the regulation of plasma VWF levels. The overall aim of this project is to explore the role of sequence variants at genetic loci other than the VWF gene as contributors to low VWF pathological states. Three specific aims will be pursued in this project. In each of these aims, we will use genomic information derived from sequencing of a large patient population with type 1, type 3 and “Low VWF” levels in whom prior analysis has failed to identify pathogenic sequence variants. In Aim #1, genetic variability in the 5' upstream region of the VWF locus will be examined to determine the influence on VWF transcriptional activation. Two areas of the VWF regulatory region will be examined: the proximal promoter, where cis-acting elements interacting with the Hippo signaling pathway will be examined, and a distant super- enhancer element ~45 kb upstream of VWF. In Aim #2, we will evaluate the role of sequence variants of three genes involved in VWF synthesis, storage and secretion. Variants of STX-2, STXBP5 and TC2N will be examined in endothelial cell-based in vitro models and in vivo, in KO mouse models for these genes. Lastly, we will investigate several VWF clearance receptors (CLEC4M, Stablin-2 and SCARA5) and regulators of the endocytic process (FCHO2 and RAB5C), all of which have been implicated in the regulation of plasma VWF levels in GWAS studies. These experiments will incorporate the development of new transgenic mouse models to assess clearance phenotypes, and will explore the regions and modifications to VWF that mediate the interaction with its clearance receptors.

项目概要：项目3 遗传性出血性疾病，血管性血友病（VWD）的致病基础， 基因机制和变异的研究而VWD的第二类定性形式则是错义的结果 参与配体结合的von Willebrand因子（VWF）结构域中的取代，数量性状，类型 1和3 VWD，与整个VWF基因的许多不同序列变体相关。然而，在这方面， 在>500例1型和>100例3型VWD中对VWF编码区和剪接位点进行详细基因分型后， 在20例病例中，分别有约35%和约15%的患者未发现致病性变异。这 证据，沿着来自大型全基因组关联研究（GWAS）的重复结果，强烈支持 除VWF外，其他基因也参与了血浆VWF水平的调节。其总体目标是 该项目旨在探索VWF基因以外的遗传位点的序列变异作为VWF基因的贡献者的作用。 低VWF病理状态。该项目将追求三个具体目标。在每一个目标中，我们将 使用从具有1型、3型和“低”的大患者群体的测序中获得的基因组信息， 先前分析未能确定致病序列变异的VWF水平。在目标#1中，遗传 将检查VWF基因座5 ′上游区域的变异性以确定对VWF的影响 转录激活将检查VWF调节区的两个区域：近端启动子， 在那里，将检查与Hippo信号通路相互作用的顺式作用元件，以及一个遥远的超- VWF上游约45 kb的增强子元件。在目标#2中，我们将评估三个基因的序列变体的作用。 参与VWF合成、储存和分泌的基因。STX-2、STXBP 5和TC 2N的变体将在 在基于内皮细胞的体外模型和体内KO小鼠模型中检查这些基因。最后， 我们将研究几种VWF清除受体（CLEC 4 M，Stablin-2和SCARA 5）和VWF清除的调节因子。 内吞过程（FCHO 2和RAB 5C），所有这些都涉及血浆VWF的调节 GWAS研究中的水平。这些实验将纳入新的转基因小鼠模型的开发 以评估清除表型，并将探索区域和修饰VWF介导的 与其清除受体相互作用。