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

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要：项目3 遗传出血疾病的致病基础von Willebrand疾病（VWD）代表了频谱遗传机制和变体的。虽然定性2种形式的VWD是错义的结果涉及配体结合的von Willebrand因子（VWF）领域的替换，定量性状，类型 1和3 VWD与整个VWF基因的许多不同序列变体有关。然而，以下> 500类型1和> 100型3 VWD的VWF编码区域和香料站点的详细基因分型病例分别在约35％和约15％的患者中尚未确定致病性变异。这证据，以及大型基因组范围研究（GWAS）的重复结果，强烈支持除VWF外，基因的参与参与了血浆VWF水平的调节。总体目的项目是探索序列变体在遗传位置的作用，而不是VWF基因作为贡献者 VWF病理状态低。该项目将追求三个具体目标。在每个目标中，我们都会使用根据1型，3型和“低的患者人群进行测序得出的基因组信息 VWF”级别，其中先前的分析未能识别病原序列变体。在AIM＃1中将检查VWF基因座的5'上游区域的可变性，以确定对VWF的影响转录激活。 VWF监管区域的两个区域将进行检查：代理启动子，其中将检查与河马信号通路相互作用的顺式作用元素，并进行遥远的超级 - 增强剂元素〜VWF上游〜45 kb。在AIM＃2中，我们将评估三个序列变体的作用涉及VWF合成，存储和分泌的基因。 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检查的这些基因的KO小鼠模型。最后，我们将调查几个VWF清除接收器（CLEC4M，Stablin-2和Scara5）和监管机构内吞过程（FCHO2和RAB5C），所有这些都暗示了等离子体VWF的调节 GWAS研究的水平。这些实验将结合新的转基因小鼠模型的发展评估清除表型，并将探索介导的区域和修改与清除接收器的互动。