The Molecular Genetics of Hemostasis

项目总结 这项建议将继续这一研究计划在3个相关领域的长期重点：1) 血管性血友病因子(VWF)功能紊乱的分子发病机制，2)遗传因素 其他遗传性出血和凝血疾病的表现，以及3)蛋白质运输的调节 从内质网到高尔基体及其在血液疾病发病机制中的作用。VWF是一个关键 凝血系统的组成部分，其缺陷会导致最常见的遗传性出血 人类疾病，von Willebrand病(VWD)。VWF水平升高是高血压的主要危险因素 血栓形成，失去ADAMTS13对VWF的处理导致血栓性血小板减少性紫癜 (TTP)。该项目将利用基因组技术的最新变革性进展来揭示小说 有助于控制VWF和ADAMTS13功能的途径，并为 医学“的方法来治疗这些疾病。一个先前定位于人类的新的vwf调控基因 2号染色体将通过基因组序列分析在另外一个大的人类队列中被识别 通过在实验室小鼠中进行“基因组编辑”的建模来探索受试者及其功能。类似的工具将 用于鉴定定位于小鼠5号染色体上的一个新的TTP易感性修饰基因。我们将 还要收集所有可能的单一氨基酸替代对功能影响的综合数据集 在VWF A1和A2结构域内提供潜在人类突变的完整清单 2A、2M和2B VWD。这些数据将解决日益重要的临床问题“变种 不确定的意义“，这是整个人类遗传学领域的一个关键挑战，应该为 仅根据DNA序列即可对VWD进行最终诊断和细分，从而实现真正的精确度 医学“，并为其他遗传病提供了一个有用的范例。本节目还将重点关注 确定与静脉血栓栓塞症(VTE)疾病易感性有关的新基因，这两种基因都是通过直接 人类静脉血栓栓塞症患者的基因组序列分析，以及广泛的全基因组突变筛选 血栓抑制基因在实验室小鼠中的应用。最后，在“床边”到“长凳”的翻译中，实验室有 扩大了对罕见的遗传性出血性疾病的研究，即第五和第八因子联合缺乏，以 探索细胞运输通路的基本功能导致对分子的意想不到的见解 先天性红细胞生成性贫血II的发病机制和血浆胆固醇水平的调节。这些 研究结果现在正从“替补席”回到“床边”，有可能提供更好的 相关疾病的诊断和治疗。总而言之，这项研究计划将应用尖端技术 基因和基因组技术，以确定改变许多疾病的风险和严重性的关键基因 血液和心脏疾病，以及关于可能发生的基本生物过程的信息 为未来诊断和治疗这些疾病的新方法奠定了基础。

PROJECT SUMMARY This proposal will continue the longstanding focus of this research program in 3 related areas: 1) the molecular pathogenesis of disorders in von Willebrand factor (VWF) function, 2) the genetic factors that modify the manifestations of other inherited bleeding and blood clotting diseases, and 3) regulation of protein transport from the ER to the Golgi apparatus and its role in the pathogenesis of blood diseases. VWF is a key component of the blood coagulation system whose deficiency resulting in the most common inherited bleeding disorder in humans, von Willebrand disease (VWD). Elevated levels of VWF are a major risk factor for thrombosis, with loss of VWF processing by ADAMTS13 resulting in thrombotic thrombocytopenic purpura (TTP). This project will exploit recent transformative advances in genomic technology to uncover novel pathways contributing to the control of VWF and ADAMTS13 function and lay the foundation for a “precision medicine” approach to these disorders. A novel VWF regulatory gene previously mapped to human chromosome 2 will be identified through genomic sequence analysis in an additional large cohort of human subjects and its function explored through modeling by “genome editing” in laboratory mice. Similar tools will be used to characterize a novel modifier gene for TTP susceptibility mapped to mouse chromosome 5. We will also assemble a comprehensive dataset for the functional impact of all possible single amino acid substitutions within the VWF A1 and A2 domains to provide a complete inventory of potential human mutations causing type 2A, 2M and 2B VWD. These data will address the increasingly important clinical problem of “variant of uncertain significance”, a key challenge for the entire field of human genetics, and should lay the foundation for eventual diagnosis and subclassification of VWD on the basis of DNA sequence alone, enabling true “precision medicine”, and serving as a useful paradigm for other genetic diseases. This program will also focus on identifying novel genes that contribute to venous thromboembolic (VTE) disease susceptibility, both by direct genomic sequence analysis in human VTE patients, as well as a broad whole genome mutagenesis screen for thrombosis suppressor genes in laboratory mice. Finally, in a “bedside” to “bench” translation, the lab has broadened its studies of the rare inherited bleeding disorder, combined deficiency of factors V and VIII, to explore the basic function of cellular transport pathways leading to unexpected insights into the molecular pathogenesis of congenital dyserythropoietic anemia II and the regulation of plasma cholesterol levels. These findings are now circling back from the “bench” to the “bedside”, with the potential to provide improved diagnosis and therapy for related diseases. Taken together, this research program will apply cutting-edge genetic and genomic technologies to identify critical genes modifying the risk and severity for a number of blood and heart diseases, as well as yielding information about fundamental biologic processes that could lay the ground work for future novel approaches to the diagnosis and treatment of these disorders.