Project 4: Epigenetic Mechanisms Modulating VWF

项目 4：调节 VWF 的表观遗传机制

• 批准号: 10113379
• 负责人: Christopher Ng
• 金额: $ 30.29万
• 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/10113379
• 关键词: 5' Untranslated Regions; ABO blood group system; AVPR2 gene; Affect; Age; Aging; Binding; Binding Sites; Biological Assay; Biology; Blood; Cell model; ChIP-seq; Code; CpG Islands; Data; Endothelial Cells; Epigenetic Process; GNA12 gene; General Population; Genes; Genetic Transcription; Hematological Disease; Hemorrhage; Hemostatic Agents; Histone Acetylation; Histones; Hypermethylation; Individual; Inflammation; Interleukin-1 beta; Messenger RNA; Methylation; MicroRNAs; Modification; Mutation; Myocardial Infarction; NFAT5 protein; PF4 Gene; Pathway interactions; Patients; Pattern; Plasma; Positioning Attribute; Proteins; RNA Polymerase II; Regulation; Reporter; Reporting; Research; Risk Factors; Role; Site-Directed Mutagenesis; Small Interfering RNA; Stroke; TNF gene; Testing; Thrombosis; Transcriptional Regulation; Transfection; Variant; age effect; base; bisulfite sequencing; cytokine; differential expression; disease phenotype; epigenetic regulation; experimental study; genome wide methylation; histone methylation; histone modification; locked nucleic acid; novel; overexpression; programs; promoter; release factor; single-cell RNA sequencing; thrombotic; transcription factor; transcriptome sequencing; vector; von Willebrand Disease; von Willebrand Factor

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.

项目摘要：项目4（ESI） von willebrand因子（VWF）是一种必不可少的止血蛋白，VWF水平的改变是相关的 出血和血栓形成。 VWF水平<50 IU/DL代表出血的危险因素，而VWF级别<30 IU/DL定义了Willebrand疾病（VWD），通常与VWF基因中的突变有关。 或者，高水平的VWF与血小板状况有关，例如心肌梗塞和 中风。普通人群中VWF水平的广泛分布表明有多种因素 调节VWF水平。迄今为止，主要的修饰符，例如ABO血型和基因的特定变体 与VWF释放有关（STXBP1，GNA12）或清除（LRP1，AVPR2，ACE）仅占30-40％ 已知的变化。该提案的目的是确定调节VWF的替代机制 水平。我们的中心假设是内皮中的特定转录和表观遗传机制 细胞是VWF水平的关键决定因素。该假设基于最近的报告和我们的初步 microRNA水平（miR），转录因子和表观遗传机制的数据都可以修改VWF 水平。我们的位置是使用主要患者衍生的血液生长来检验这一假设的独特位置 精确复制疾病表型的内皮细胞（BOEC）。我们将通过 以下三个目标，（1）确定影响BOEC中VWF表达的转录机制， （2）确定VWF对VWF表达的甲基化和组蛋白 - 乙酰化状态的贡献 从BOEC和（3）确定转录和表观遗传机制在VWF水平上的作用 老化。我们的初步数据和BOEC模型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）确认VWF的新型转录调节剂 低VWF BOEC，例如miR-24和TCF4，（2）证实表观遗传修饰剂的调节作用，例如 VWF启动子甲基化和组蛋白乙酰化，在确定VWF水平时，（3）证明了新颖的 转录和表观遗传调节，可以解释衰老对VWF水平的影响。另外，我们的 未偏置的RNA测序，甲基化阵列和microRNA阵列也可能确定其他目标 VWF调节。在全球范围内，通过评估转录和表观遗传调节剂在VWF水平上的作用 预计我们将确定VWF表达和功能的新型机制。这样的结果很重要 正如他们期望的那样，他们将了解对VWF编码区域外的修饰符的理解 影响VWF水平并代表以前不好的VWF调节的新途径 检查。