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: 10304868
• 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 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10304868
• 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; 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.

概括 我们对遗传血小板功能障碍的分子基础有长期的研究兴趣。 转录因子runx1是造血，巨型摩毛虫和血小板的主要调节剂。 该项目的总体目标是获得对血小板功能障碍的分子基础的见解 与人类runx1突变以及在血小板/巨核细胞（MK）中调节的基因相关的基因 通过针对Runx1单位缺陷（RHD）患者的研究，其特征是农场血小板减少症， 血小板功能障碍和急性白血病的倾向。它们在血小板颗粒和 血小板反应受损。我们对RHD患者的血小板表达分析，这是第一个血小板之一 在遗传血小板功能障碍的患者中进行了分析研究，显示了几个基因已降低 受调节，有些是直接的runx1靶标，并且会影响血小板/MK生物学的特定方面。 RHD是一个 重要的人类模型和未开发的信息储存库中的血小板/ MK生物学。研究 由当前R01奖的支持取得了非常成功，并扩展了Runx1的相关性 从出血疾病到简历疾病的基因调节。我们提出了关于2的RUNX1调控的研究 RHD血小板中表达降低的基因：凝结因子XIIIA（Gene F13a）和 磷酸二酯酶E5A（PDE5A）;关于他们的调节知之甚少。目标1是研究机制 RHD中F13A和PDE5A的MK/血小板表达降低的影响。 F13a由MK合成， （〜3％的血小板蛋白），调节凝块回缩。 PDE5A调节CGMP级别，CGMP水平是 血小板反应。我们将在RHD患者血小板和培养细胞中进行研究（HEL细胞和MK 由RHD患者由IPSC产生）。我们将评估这些基因在 与心脏病患者的未来死亡和MI有关。 runx1从两个表示 替代启动子 - 远端P1和代理P2启动子，导致两个不同的蛋白质，Runx1c和 runx1b，具有不同的效果。 AIM 2是研究MK/血小板基因的差异调节 runx1b和runx1c。最近的证据表明，Runx1c由Runx1自动调节。目标3是 研究runx1b和runx1c对Runx1的自动调节。我们的研究将提供重要的新 在RHD中的异常血小板/MK机制中的信息，runx1的差异基因调节 变体及其下游影响以及与简历疾病的关系。他们将为 制定较新的抗血栓形成策略。

项目成果

Alterations in insulin-signaling and coagulation pathways in platelets during hyperglycemia-hyperinsulinemia in healthy non-diabetic subject.

• DOI: 10.1016/j.thromres.2014.06.029
• 发表时间: 2014-09
• 期刊: THROMBOSIS RESEARCH
• 影响因子: 7.5
• 作者: Rao, A. Koneti;Freishtat, Robert J.;Jalagadugula, Gauthami;Singh, Anamika;Mao, Guangfen;Wiles, Andrew;Cheung, Peter;Boden, Guenther
• 通讯作者: Boden, Guenther

Transcription Factor RUNX1 Regulates Platelet PCTP (Phosphatidylcholine Transfer Protein): Implications for Cardiovascular Events: Differential Effects of RUNX1 Variants.

• DOI: 10.1161/circulationaha.116.023711
• 发表时间: 2017-09-05
• 期刊: Circulation
• 影响因子: 37.8
• 作者: Mao G;Songdej N;Voora D;Goldfinger LE;Del Carpio-Cano FE;Myers RA;Rao AK
• 通讯作者: Rao AK

Platelet disorders: the next generation is in.

血小板疾病：下一代已到来。

• DOI: 10.1182/blood-2016-04-703215
• 发表时间: 2016
• 期刊: Blood
• 影响因子: 20.3
• 作者: Rao,AKoneti;Songdej,Natthapol
• 通讯作者: Songdej,Natthapol

Spotlight on FLI1, RUNX1, and platelet dysfunction.

• DOI: 10.1182/blood-2013-10-533166
• 发表时间: 2013-12
• 期刊: Blood
• 影响因子: 20.3
• 作者: A. Rao

Nuclear factor-κB regulates expression of platelet phospholipase C-β2 (PLCB2).

• DOI: 10.1160/th15-09-0749
• 发表时间: 2016-10-28
• 期刊: Thrombosis and haemostasis
• 影响因子: 6.7
• 作者: Mao G;Jin J;Kunapuli SP;Rao AK
• 通讯作者: Rao AK