The Molecular Genetics of Hemostasis

项目摘要 该提案将继续在3个相关领域的研究计划的长期重点：1） 血管性血友病因子（VWF）功能障碍的分子发病机制，2）修饰 其他遗传性出血和凝血疾病的表现，以及3）蛋白质转运的调节 从内质网到高尔基体及其在血液病发病机制中的作用。VWF是一把钥匙 凝血系统的一种成分，其缺陷导致最常见的遗传性出血 在人类中，血管性血友病（von Willebrand disease，VWD）。VWF水平升高是一个主要的危险因素， 血栓形成，伴有ADAMTS13对VWF的加工丧失，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， 有助于控制VWF和ADAMTS13功能的途径，并为“精确”治疗奠定基础。 医学”的方法来治疗这些疾病。一个新的人血管性血友病因子调控基因 将通过基因组序列分析在另外一个大的人类群体中鉴定2号染色体。 通过在实验室小鼠中进行“基因组编辑”建模，探索其主题和功能。类似工具将 用于表征定位于小鼠5号染色体的TTP易感性的新修饰基因。我们将 还为所有可能的单个氨基酸取代的功能影响组装了综合数据集 在VWF A1和A2结构域内，以提供引起型VWF的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，“变异的 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 VWD的最终诊断和亚分类仅基于DNA序列，实现真正的“精确 医学”，并作为其他遗传疾病的有用范例。该计划还将重点关注 鉴定导致静脉血栓栓塞（VTE）疾病易感性的新基因， 基因组序列分析，以及广泛的全基因组诱变筛选， 血栓抑制基因的研究。最后，在“床边”到“长凳”的翻译中，实验室已经 扩大了对罕见的遗传性出血性疾病的研究，同时缺乏因子V和VIII， 探索细胞运输途径的基本功能，从而对分子生物学产生意想不到的见解。 先天性红细胞生成不良性贫血II的发病机制和血浆胆固醇水平的调节。这些 研究结果现在正在从“板凳”回到“床边”，有可能提供改善的 相关疾病的诊断和治疗。总的来说，这项研究计划将应用尖端的 基因和基因组技术，以确定关键基因改变的风险和严重性，为一些 血液和心脏疾病，以及产生有关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.