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.

Zimmerman VWD 生物学计划 (ZPB-VWD) 的项目 1 重点关注分子影响 VWF 对临床 VWD 的影响。它整合了两个现有队列 – 1) 之前的回顾性 PPG 队列 在齐默尔曼 VWD 分子和临床生物学项目中诊断出来，并且 2) 前瞻性 R01 队列新兵。目标 1 对 1、2 和 3 型 VWD 受试者纵向进行实验室研究 以及 VWF 水平降低的群体，我们统称为低冯维勒布兰德因子，其中包括 那些之前可能被标记为“轻度 VWD”且 VWF <正常但出血很少或没有出血的患者。 这些可能被认为是出血的危险因素，但不是疾病。我们不仅要学习 实验室表型随时间的变化，但也可以通过临时出血评估来研究它们 自 BAT 评估出血以来，通过评分 (iBAT) 半定量评估“新的临床出血”量 但分数已饱和，无法量化临时出血。此外，目标 1 将量化评估 与新基质蛋白（例如肌球蛋白）或新功能受体（例如血小板 GP）的潜在异常结合 IIb-IIIa (2b3a)。在目标 2 中，将研究 2 型功能变异以确定诊断的保真度（类型 2A、2B、2M 和 2N) 本地表型分配与集中分配的表型分配之间的差异。动物模型 2 型变体已经/将要开发并使用出血和血栓形成测试进行比较（尾部 出血、模板出血、VenaFlux 和激光损伤评估。目标 3 将包括我们的科目 合并患有 1 型或 3 型 VWD 或 LVWF 的队列，并在这些现有队列中添加现有队列，类似地 定义，来自金斯顿和都柏林的队列，但没有 VWF 突变，并且阵列比较结果为阴性 基因组杂交 (aCGH) 或多重连接依赖性探针扩增 (MLPA)。在我们的 队列中，我们至少有 3 名 3 型 VWD 受试者，没有 VWF 序列变异，aCGH 呈阴性，并且 没有抗 VWF 抗体。此外，我们还有 >40 个 1 型 VWD 和 >180 个 LVWF 受试者样本 NSV 和负 aCGH。因此，我们将首先通过全基因组研究这些 1 型和 3 型受试者 在 Core B 中使用 NGS 测序进行测序 (WGS)。当识别出单个候选基因时， 将对受影响的 (AFM) 和未受影响的家庭成员 (UFM) 进行测序以确定 与表型的联系。然后，候选 SV 将根据项目 1-4 是否符合条件而被分包给项目 1-4。 1）VWF内含子变化（项目1）； 2）涉及碳水化合物或碳水化合物受体（项目2）； 3） VWF 编码区之外的机制（项目 3）；或 4) 提出表观遗传机制（项目 4）。 对于可能被识别的常见或科学上独特的原因，将在以下环境中制作这些动物模型 核心 C 并在各种项目中进行了研究。通过项目 1，我们希望深入了解这种疾病 导致 VWD 的机制以及它们如何通过发育的动物影响实验室和临床表型 模型，并协助确定 1 型和 3 型 VWD 的外源原因。