Project 1: Molecular Impact of VWF on Clinical VWD

项目 1：VWF 对临床 VWD 的分子影响

• 批准号: 10113376
• 负责人: ROBERT R MONTGOMERY
• 金额: $ 33.35万
• 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/10113376
• 关键词: 1 year old; Acute; Adult; Affect; Age; Animal Model; Antibodies; Assessment tool; Binding; Biological Assay; Biology; Blood Platelets; Candidate Disease Gene; Carbohydrates; Cells; Child; Childbirth; Clinical; Clinical assessments; Code; Collagen; DNA; DNA Sequencing Facility; Databases; Defect; Diagnosis; Disease; Elderly; Engineered Gene; Enhancers; Enrollment; Epigenetic Process; F8 gene; Family member; Farming environment; Foundations; Frequencies; Funding; Genes; Genetic; Genomics; Genotype; Hemophilia A; Hemorrhage; Human; Individual; Introns; Label; Laboratories; Laboratory Study; Laser injury; Ligation; Modeling; Modification; Molecular; Mutation; Myosin ATPase; Newly Diagnosed; Operative Surgical Procedures; Pathway interactions; Patients; Phenotype; Physiological; Plasma; Platelet Membrane Glycoprotein IIb; Process; Program Research Project Grants; Proteins; Rattus; Research Design; Risk Factors; Rodent Model; Sampling; Severities; Stress; Symptoms; Tail; Testing; Thrombocytopenia; Thrombosis; Time; United States National Institutes of Health; VWF gene; Variant; Visit; age related; base; biobank; carbohydrate receptor; clinical phenotype; cohort; comparative genomic hybridization; follow-up; genome sequencing; head-to-head comparison; improved; in vivo; insight; mouse model; programs; promoter; prospective; receptor; recruit; reduce symptoms; thrombotic; treatment center; von Willebrand Factor; whole genome

Project 1 in the Zimmerman Program for the Biology of VWD (ZPB-VWD) is focused on the Molecular Impact of VWF on Clinical VWD. It consolidates the two existing cohorts – 1) retrospective PPG cohort previously diagnosed in the Zimmerman Program for the Molecular and Clinical Biology of VWD and 2) the prospective R01 cohort recruits. In Aim 1 laboratory studies are done longitudinally on the Type 1, 2, and 3 VWD subjects as well as the group with reduced VWF levels that we collectively call Low von Willebrand Factor that includes those that might have been previously labeled as “Mild VWD” with VWF <normal but little or no bleeding. These may be considered as having a risk factor for bleeding but not a disease. Not only will we study the changes with time of their laboratory phenotype but also study them with and interim Bleeding Assessment Score (iBAT) to semiquantitatively assess the amount of “new clinical bleeding” since BATs assess bleeding but the score saturates and cannot quantify interim bleeding. Additionally, Aim 1 will quantifiably evaluate potential abnormal binding to new matrix proteins (e.g. myosin) or new functional receptors (e.g. platelet GP IIb-IIIa (2b3a). In Aim 2 type 2 functional variants will be studied to determine the fidelity of diagnosis (types 2A, 2B, 2M, and 2N) between phenotypic assignments locally to those centrally assigned. Animal models of type 2 variants have been/will be developed and compared using bleeding and thrombosis testing (tail bleeding, template bleeding, VenaFlux, and Laser-injury assessment. Aim 3 will include subjects in our combined cohort who have type 1 or 3 VWD or LVWF and to these existing cohorts add existing, similarly defined, cohort from Kingston and Dublin but have no VWF mutation and are negative by Array Comparative Genome Hybridization (aCGH) or Multiplex Ligation-dependent Probe Amplification (MLPA). Within our cohort, we have at least 3 type 3 VWD subjects with no VWF sequence variant, are negative by aCGH, and have no anti-VWF antibodies. In addition we have >40 type 1 VWD and >180 LVWF subject samples with NSV and negative aCGH. Therefore we will first study these type 1 and 3 subjects by whole Genome Sequencing (WGS) using NGS sequencing in Core B. When individual candidate genes are identified, sequencing will be performed on the affected (AFM) and unaffected family members (UFM) to ascertain linkage with phenotype. Candidate SV will then be farmed out to Projects 1-4, based upon whether they are 1) VWF intronic changes (Project 1); 2) involve carbohydrates or carbohydrate receptors (Project 2); 3) mechanisms outside the VWF coding region (Project 3); or 4) suggest an epigenetic mechanism (Project 4). For frequent or scientifically unique causes that might be identified, animal models of these will be produced in Core C and studied in the various projects. Through Project 1 we expect to gain insight into the disease mechanisms causing VWD and how they affect laboratory and clinical phenotypes through developed animal models, and to assist with determining extragenic causes of both type 1 and type 3 VWD.

VWD生物学（ZPB-VWD）的Zimmerman计划中的项目1侧重于分子影响 VWF的临床VWD。它巩固了两个现有队列 - 1）以前回顾PPG队列 在VWD的分子和临床生物学的Zimmerman计划中被诊断为2） R01队列新兵。在AIM 1实验室研究中，对1、2和3型VWD受试者进行了纵向进行 以及具有降低的VWF级别的小组，我们将其称为低von Willebrand因素，其中包括 那些可能以前被标记为具有VWF <正常或没有出血的“轻度VWD”的人。 这些可能被认为是出血的危险因素，而不是疾病。我们不仅会研究 随着实验室表型的时间而变化，但也通过临时出血评估进行研究 分数（IBAT）半定量评估“新临床出血”的量，因为蝙蝠评估出血 但是得分饱和，无法量化临时出血。此外，AIM 1将量化评估 潜在的异常结合与新基质蛋白（例如肌球蛋白）或新功能受体（例如血小板GP） IIB-IIIA（2B3A）。在AIM 2中，将研究2型功能变体，以确定诊断的保真度（类型 2a，2b，2m和2n）在本地分配的表型分配之间。动物模型 使用出血和血栓形成测试（尾巴）已开发和比较2型变体（尾巴） 出血，模板出血，venaflux和激光伤害评估。 AIM 3将包括我们的主题 具有1型或3型VWD或LVWF的组合队列，并向这些现有同类群体增加现有的，同样 定义的，来自金斯敦和都柏林的队列，但没有VWF突变，并且对数组的比较是负的 基因组杂交（ACGH）或多重连接依赖性探针扩增（MLPA）。在我们内部 队列，我们​​至少有3个3型VWD受试者，没有VWF序列变体，ACGH为负，并且 没有抗VWF抗体。此外，我们有> 40型1型VWD和> 180 lvwf主题样品 NSV和负ACGH。因此，我们将首先根据整个基因组研究这些1和3的主题 使用核心B中的NGS测序进行测序（WGS）。确定单个候选基因时， 测序将对受影响的（AFM）和未受影响的家庭成员（UFM）进行确定 与表型的联系。然后，候选人SV将根据他们是否是 1）VWF内含子更改（项目1）； 2）涉及碳氢化物或碳水化合物受体（项目2）； 3） VWF编码区域以外的机制（项目3）；或4）提出一种表观遗传机制（项目4）。 对于可能被识别的经常或科学独特的原因，将在其中产生这些动物模型 核心C和Studiod在各种项目中。通过项目1，我们希望能深入了解该疾病 导致VWD及其如何影响实验室和临床表型的机制 模型，并协助确定1型和3型VWD的基本原因。