Human Platelet Defects in Transcription Factor RUNX1 Haplodeficiency

转录因子 RUNX1 单倍体缺陷中的人血小板缺陷

• 批准号: 10083753
• 负责人: Angara Koneti Rao
• 金额: $ 45.4万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-18 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10083753
• 关键词: 12-HETE; Abnormal Platelet; Acute leukemia; Address; Affect; Affinity; Alpha Granule; Amino Acids; Aspirin; Award; Binding; Biology; Blood; Blood Cells; Blood Coagulation Disorders; Blood Coagulation Factor; Blood Platelets; Cardiovascular Diseases; Cell physiology; Cells; Cellular biology; Cessation of life; Clot retraction; Coagulation Process; Collaborations; Complex; Cultured Cells; Cyclic AMP; Cyclic GMP; Cytoplasmic Granules; DNA Binding; Defect; Disease; Distal; Down-Regulation; Event; Expression Profiling; Factor XIII; Factor XIIIa; Functional disorder; Future; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Heart Diseases; Hela Cells; Hematopoiesis; Hemorrhage; Homeostasis; Human; Impairment; Individual; Inherited; Injury; Link; Megakaryocytes; Megakaryocytopoieses; Molecular; Molecular Biology; Mutation; Myeloid Cells; Myocardial Infarction; N-terminal; Patients; Phosphorylation; Plasma; Platelet aggregation; Play; Predisposition; Production; Protein Isoforms; Protein Kinase C; Proteins; RUNX1 gene; Regulation; Research; Role; Site; Symptoms; T-Lymphocyte; Thrombocytopenia; Thrombopoiesis; Transcript; Transglutaminases; Universities; Variant; cardiovascular disorder risk; cardiovascular disorder therapy; clinical practice; cohort; human model; in vivo; inhibitor/antagonist; insight; interest; novel therapeutics; phosphoric diester hydrolase; promoter; response; stem cells; transcription factor

Summary We have a longstanding research interest into the molecular basis of inherited platelet dysfunction. Transcription factor RUNX1 is a master regulator of hematopoiesis, megakaryopoiesis and thrombopoiesis. The overall goal of this project is to obtain insights into the molecular basis of the platelet dysfunction associated with human RUNX1 mutations and into the genes regulated by it in platelets/megakaryocytes (MK) through studies in patients with RUNX1 haplodeficiency (RHD), characterized by familial thrombocytopenia, platelet dysfunction and a predisposition to acute leukemia. They have abnormalities in platelet granules and impaired platelet responses. Our platelet expression profiling of a RHD patient, one of the first platelet profiling studies in a patient with inherited platelet dysfunction, showed several genes are down regulated, some are direct RUNX1 targets, and affect specific aspects of platelet/MK biology. RHD is an important human model and untapped reservoir of information into platelet/ MK biology. Studies supported by the current R01 Award have been highly successful and extend the relevance of RUNX1 regulation of genes from bleeding disorder to CV disease. We propose studies on the RUNX1 regulation of 2 genes whose expression is decreased in RHD platelets: coagulation factor XIIIa (gene F13A) and phosphodiesterase E5A (PDE5A); little is known regarding their regulation. Aim 1 is to study the mechanisms and effects of decreased MK/platelet expression of F13A and PDE5A in RHD. F13A is synthesized by MK, (~3% of platelet protein) and regulates clot retraction. PDE5A regulates cGMP levels, a major regulator of platelet responses. We will perform studies in RHD patient platelets and in cultured cells, (HEL cells and MK generated from IPSCs from a RHD patient). We will assess association of RUNX1 regulation of these genes in vivo and in relation to future death and MI in patients with heart disease. RUNX1 is expressed from two alternate promoters – a distal P1 and a proximal P2 promoter, leading to two distinct proteins, RUNX1C and RUNX1B, with differential effects. Aim 2 is to study the differential regulation of MK/platelet genes by the RUNX1B and RUNX1C. Recent evidence suggests that RUNX1C is autoregulated by RUNX1. Aim 3 is to study the autoregulation of RUNX1 by RUNX1B and RUNX1C. Our studies will provide important, new information into the aberrant platelet/MK mechanisms in RHD, the differential gene regulation by RUNX1 variants and their downstream effects, and the relationship to CV disease. They will lay the basis for developing newer antithrombotic strategies.

总结 我们对遗传性血小板功能障碍的分子基础有着长期的研究兴趣。 转录因子RUNX 1是造血、巨核细胞生成和血小板生成的主要调节因子。 本项目的总体目标是深入了解血小板功能障碍的分子基础 与人RUNX 1突变相关，并进入血小板/巨核细胞（MK）中受其调节的基因 通过对RUNX 1单倍缺陷（RHD）患者的研究，其特征在于家族性血小板减少症， 血小板功能障碍和易患急性白血病他们的血小板颗粒异常， 血小板反应受损。我们对一名风湿性心脏病患者的血小板表达谱进行了分析， 对一名患有遗传性血小板功能障碍的患者进行的分析研究显示， 受调节，一些是直接RUNX 1靶点，并影响血小板/MK生物学的特定方面。RHD是一个 重要人类模型和血小板/ MK生物学中未开发的信息库。研究 在当前R 01奖的支持下，取得了巨大成功，并扩展了RUNX 1的相关性 从出血性疾病到CV疾病的基因调控。我们建议研究RUNX 1对2 在RHD血小板中表达降低的基因：凝血因子XIIIa（基因F13 A）和 磷酸二酯酶E5 A（PDE 5A）;关于其调节知之甚少。目的一是研究其作用机制 以及MK/血小板F13 A和PDE 5A表达降低在RHD中的作用。F13 A由MK合成， （约3%的血小板蛋白），并调节凝块收缩。PDE 5A调节cGMP水平，这是一种主要的调节剂， 血小板反应我们将在风湿性心脏病患者血小板和培养细胞（HEL细胞和MK）中进行研究 由来自RHD患者的IPSC产生）。我们将评估RUNX 1对这些基因的调控在 在心脏病患者中，与未来死亡和MI相关。RUNX 1由两个 交替启动子-远端P1和近端P2启动子，导致两种不同的蛋白质，RUNX 1C和 RUNX 1B，具有差异效应。目的二是研究血小板膜蛋白对MK/血小板基因的差异调控。 RUNX 1B和RUNX 1C。最近的证据表明，RUNX 1C是由RUNX 1自动调节的。目标3是 研究RUNX 1B和RUNX 1C对RUNX 1的自身调节作用。我们的研究将提供重要的，新的 RHD中异常血小板/MK机制的信息，RUNX 1的差异基因调控 变异及其下游效应，以及与CV疾病的关系。他们将为 开发新的抗血栓策略。