Project 4: Epigenetic Mechanisms Modulating VWF

项目摘要：项目4(ESI) Von Willebrand因子(Vwf)是一种重要的止血蛋白，vwf水平的改变与vwf有关。 有出血和血栓形成。VWF水平&lt；50IU/dL代表出血的危险因素，而VWF水平&lt；30 Iu/dL定义为威勒布兰德病(VWD)，通常与VWF基因突变有关。 或者，高水平的VWF与血栓形成有关，如心肌梗死和 卒中。VWF水平在普通人群中的广泛分布表明存在多种因素 来调节VWF水平。到目前为止，主要的修饰因素，如ABO血型和基因的特定变异 与VWF释放(STXBP1、GNA12)或清除(LRP1、AVPR2、ACE)有关的仅占30%-40% 已知的变种。这项提案的目标是确定监管VWF的替代机制 级别。我们的中心假设是内皮细胞中特定的转录和表观遗传机制 细胞是VWF水平的关键决定因素。这一假设是基于最近的报道和我们初步的 有数据表明，microRNAs水平(MiRs)、转录因子和表观遗传机制都可以改变vWF 级别。我们处于独特的地位，可以使用我们的原始患者来源的血液培养来检验这一假设 准确复制疾病表型的内皮细胞(BOECs)。我们将通过以下方式测试我们的假设 根据以下三个目标，(1)确定影响BOEC中VWF表达的转录机制， (2)VWF甲基化和组蛋白乙酰化状态对VWF表达的影响 来自BOECs和(3)转录和表观遗传机制在VWF水平上的作用 衰老。我们的初步数据和我们在BOEC模型方面的专业知识使我们能够很好地执行这些建议 研究目的。这些目标的成功完成将(1)证实VWF在 低VWF BOEC，如miR-24和TCF4，(2)确认表观遗传修饰物的调节作用，如 VWF启动子甲基化和组蛋白乙酰化在确定VWF水平中的作用，以及(3)展示了新的 转录和表观遗传调控可能解释了衰老对VWF水平的影响。此外，我们的 无偏见的RNA测序、甲基化阵列和微RNA阵列也可能识别更多的靶点 VWF法规。在全球范围内，通过评估转录和表观遗传调节因子在VWF水平上的作用，它是 预计我们将发现VWF表达和功能的新机制。这样的结果意义重大 因为它们有望促进对VWF编码区之外的修饰物如何能够 影响VWF水平，并代表以前不太好的VWF调节的新途径 检查过了。

PROJECT SUMMARY: PROJECT 4 (ESI) Von Willebrand factor (VWF) is an essential hemostatic protein and alterations of VWF levels are associated with bleeding and thrombosis. VWF levels < 50 IU/dL represent a risk factor for bleeding while VWF levels < 30 IU/dL define Willebrand disease (VWD) and are often associated with mutations in the VWF gene. Alternatively, high levels of VWF are associated with thrombotic conditions such as myocardial infarction and stroke. The wide distribution of VWF levels in the general population indicates that there are multiple factors that regulate VWF levels. To date, major modifiers such as the ABO blood group and specific variants in genes implicated in VWF release (STXBP1, GNA12) or clearance (LRP1, AVPR2, ACE) account for only 30-40% of the known variation. The objective of this proposal is to identify alternative mechanisms that regulate VWF levels. Our central hypothesis is that specific transcriptional and epigenetic mechanisms in endothelial cells are critical determinants of VWF levels. The hypothesis is based on recent reports and our preliminary data that microRNAs levels (miRs), transcription factors, and epigenetic mechanisms can all modify VWF levels. We are uniquely positioned to test this hypothesis using our primary patient-derived blood outgrowth endothelial cells (BOECs) that replicate accurately the disease phenotype. We will test our hypothesis via the following three aims, (1) Determination of transcriptional mechanisms that affect VWF expression in BOECs, (2) Determination of the contribution of methylation and histone-acetylation status of VWF on VWF expression from BOECs and (3) Determination of the role of transcriptional and epigenetic mechanisms on VWF levels in aging. Our preliminary data and our expertise in BOEC models positions us well to carry out these proposed research aims. Successful completion of these aims will (1) confirm novel transcriptional regulators of VWF in low VWF BOECs, such as miR-24 and TCF4, (2) confirm the regulatory role of epigenetic modifiers, such as VWF promoter methylation and histone acetylation, in determining VWF levels, and (3) demonstrate novel transcriptional and epigenetic regulation that may explain the effects of aging on VWF levels. In addition, our un-biased RNA sequencing, methylation arrays, and microRNA arrays may also identify additional targets for VWF regulation. Globally, by evaluating the role of transcriptional and epigenetic regulators on VWF levels it is anticipated that we will identify novel mechanisms of VWF expression and function. Such results are significant as they are expected to advance the understanding of how modifiers outside of the VWF coding region can influence VWF levels and represent novel pathways of VWF regulation that have previously not been well examined.