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: 10584537
• 负责人: DAVID P LILLICRAP
• 金额: $ 26.05万
• 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 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10584537
• 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; Heterozygote; 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; 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.

项目总结：项目3 遗传性出血性疾病von Willebrand病(Vwd)的致病基础代表了一种谱系。 遗传机制和变异。而VWD的定性类型2是误解的结果 与配基结合有关的von Willebrand因子(VWF)结构域上的替换、数量性状、类型 1和3VWD与VWF基因中许多不同的序列变异有关。然而， 在对&gt；500 1型和&gt；100型3型vwd的vwf编码区和剪接部位进行详细的基因分型后 2例中，分别有~35%和~15%的患者未发现致病变异。这 证据，以及来自大基因组全关联研究(GWAS)的重复结果，有力地支持 除VWF外，基因还参与调节血浆VWF水平。这样做的总体目标是 该项目是探索除vwf基因以外的遗传基因上的序列变异作为 低VWF病理状态。这个项目将追求三个具体目标。在每个目标中，我们都将 使用通过对大量1型、3型和“低”患者进行测序而获得的基因组信息 VWF“水平，在此之前的分析未能确定致病序列变异。在目标1中，遗传 将检查VWF基因5‘上游区域的可变性，以确定对VWF的影响 转录激活。我们将研究VWF调控区域的两个区域：近端启动子， 在那里将检查与河马信号通路相互作用的顺式作用元件，以及遥远的超级- VWF上游约45kb的增强子元件。在目标2中，我们将评估三个序列变体的作用 参与VWF合成、储存和分泌的基因。STX-2、STXBP5和TC2N的变体将是 在基于内皮细胞的体外模型和体内模型中，在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(CLEC4M、Sablin-2和SCARA5)及其调节因子 内吞过程(FCHO2和RAB5C)，所有这些都与血浆VWF的调节有关 全球气候变化研究中的水平。这些实验将包括开发新的转基因小鼠模型。 评估清除表型，并将探索介导VWF的区域和修饰 与其清除受体相互作用。