Platelet TSP1 Mediates Vascular Disease and pulmonary hypertension in Sickle Cell

血小板 TSP1 介导镰状细胞中的血管疾病和肺动脉高压

• 批准号: 8849490
• 负责人: Enrico M Novelli
• 金额: $ 13.22万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8849490
• 关键词: Adhesions; Adult; Alpha Granule; Animals; Area; Automobile Driving; Award; Benign; Binding; Biological Availability; Biology; Biometry; Blocking Antibodies; Blood Platelets; Blood Vessels; Bone Marrow; CD47 gene; Cell Culture System; Cell Surface Receptors; Chimera organism; Chronic; Clinical; Clinical Research; Clinical Trials; Clinical Trials Design; Complement; Cutaneous; Data; Development; Disease; Disease Marker; Endothelial Cells; Endothelin; Endothelin A Receptor; Endothelin-1; Endothelium; Epidemiology; Equilibrium; Erythrocytes; Event; Functional disorder; Funding; Funding Mechanisms; Gap Junctions; Generations; Genes; Genetic; Globin; Grant; Hematologist; Hematology; Hemoglobin; Hemolysis; Hemolytic Anemia; Hemostatic Agents; Hemostatic function; Histologic; Human; In Situ; In Vitro; Incubated; Infarction; Inflammatory; Institute of Medicine (U.S.); International; Investigation; Iron Overload; Kidney Failure; Knock-out; Knowledge; Laboratories; Lead; Leg; Ligands; Link; Lung; Master of Science; Mediating; Medicine; Mentored Patient-Oriented Research Career Development Award; Mentors; Methodology; Modeling; Multivariate Analysis; Mus; Mutate; Nitric Oxide; Oligonucleotides; Oxidative Stress; Oxygen; Pathogenesis; Pathology; Pathway interactions; Patients; Peer Review; Phenotype; Physiological; Plasma; Platelet Activation; Polymers; Prevention; Process; Production; Proteins; Publications; Pulmonary Hypertension; Pulmonary artery structure; Rare Diseases; Reactive Oxygen Species; Research; Research Activity; Resources; Risk Assessment; Risk Factors; Role; Schedule; Secondary to; Severities; Sickle Cell; Sickle Cell Anemia; Sickle Hemoglobin; Signal Pathway; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Solubility; Stream; System; Testing; Therapeutic; Thromboembolism; Thrombosis; Thrombospondin 1; Time; Training Activity; Transgenic Model; Transgenic Organisms; Translational Research; Translations; Transplantation; Treatment Efficacy; Ulcer; Universities; Vascular Diseases; Vascular remodeling; Vasoconstrictor Agents; Vasodilator Agents; Walking; Work; Writing; base; bench to bedside; career; cohort; endothelial dysfunction; experience; functional status; hemodynamics; human disease; human subject; improved; indexing; insight; interest; meetings; metabolomics; mouse model; multidisciplinary; mutant; new therapeutic target; novel; novel therapeutic intervention; patient registry; pre-clinical; preclinical study; prevent; professor; programs; receptor; receptor expression; research study; response; responsible research conduct; sickling; skills; tool; treatment duration; volunteer

DESCRIPTION (provided by applicant): This is an application for a K23 award for Dr. Enrico Novelli, a Hematologist, Assistant Professor in the Tenure Stream in the Department of Medicine and Director of the University of Pittsburgh Adult Sickle Cell Anemia Program. Dr. Novelli is establishing himself as an academic leader in the field of benign hematology with a specific focus on translational research in sickle cell disease and hemostasis. Dr. Novelli has prior extensive research experiences in hematology and has been successful in obtaining research awards for his studies on SCD. He has assembled a multidisciplinary team of mentors and advisors to guide his career towards independence and to assist with the completion of the research proposed in this application. This grant will be critical to achieve the following career objectives: (1) to develop expertise in laboratory methodologies crucial to the conduct of translational research in SCD; (2) to gain further in-depth knowledge of clinical trial design and biostatistics fundamental in the investigation of human disease; and (3) to become an expert in the vascular biology of SCD. He plans to complement his research activity with the completion of a Master of Science in Clinical Research sponsored by the University of Pittsburgh, with participation in scheduled training activities on grant writing and responsible conduct of research, and with attendance to international meetings in his areas of interest. By the end of the funding period of this K23 award, he will have several first-authored peer-reviewed publications and a critical mass of data that will form the core of an RO1 or other equivalent grant. The focus of this application is on the vascular biology of SCD. In SCD, mutant hemoglobin S polymerizes when deoxygenated, driving red blood cell (RBC)-dependent vaso-occlusion and hemolysis. These processes lead to platelet and hemostatic activation, pulmonary hypertension and vascular disease. Transgenic-knockout sickle (BERK) mice that express exclusively human ¿- and ?S-globins mimic SCD in humans by displaying reduced NO bioavailability, impaired NO-mediated vascular reactivity and pulmonary hypertension. Recently, the platelet ¿-granule protein thrombospondin-1 (TSP1) was found to potently inhibits physiologic NO signaling, via binding to the cell surface receptor CD47. Preliminary data from Dr. Novelli now demonstrate that circulating TSP1 levels are increased in the plasma of patients with SCD. Preclinical studies supporting this proposal also show that the specific TSP1-CD47 ligand receptor interaction not only inhibits NO signaling in endothelial cells, but also increases reactive oxygen species (ROS) production and endothelin A (ETA) receptor expression in smooth muscle, both canonical vasoconstrictive and mitogenic pathological signaling pathways contributing to pulmonary hypertension. Extensive preliminary data using chimera cross transplantation systems confirm that CD47 is critical in the development of PH in the transgenic sickle cell mouse. These findings inform the grant's overarching hypothesis that during platelet activation in patients with SCD, increases in circulating plasma levels of TSP1, via binding to the CD47 receptor, disrupt pulmonary vascular endothelial NO production and stimulate smooth muscle ROS production and endothelin 1 signaling, which lead to pulmonary hypertension in murine models and human subjects with SCD. This grant will also explore the hypothesis that therapeutic disruption of the TSP1-CD47 ligand-receptor interaction will both prevent and reverse pulmonary hypertension in SCD. This hypothesis will be tested via the following aims: (1) to examine for the first time whether plasma from human subjects with SCD TSP1 levels in the plasma of patients with SCD disrupt the vasodilator/vasoconstrictor balance in endothelium and smooth muscle by inhibition of NO signaling and increasing ROS and ETA levels; (2) to employ unique mutant murine models to determine the role of circulating TSP1 and CD47 in SCD-associated vasculopathy and pulmonary hypertension, and test the therapeutic efficacy of blocking TSP1-CD47 to prevent/mitigate SCD-based pulmonary hypertension; and (3) to test TSP1 levels in several large cohorts of patients with SCD and explore the correlation of TSP1 with platelet activation, pulmonary hypertension and vascular disease. This proposal is strengthened by the complementary and rich tool set used to investigate the hypothesis, the vast resources available within the Vascular Medicine Institute, a state of the art facility within the University of Pittsburgh devoted to the study of vascular biology, and captures the pioneering work of Dr. Novelli's mentors in defining hemolysis in the pathophysiology of SCD (Dr. Mark Gladwin's lab) and the TSP1-CD47 nexus as an upstream regulator of NO signaling (Dr. Jeff Isenberg's lab). Definition of this pathway will therefore present a novel therapeutic target for the vascular complications of SCD.

描述(由申请人提供)：这是对Enrico Novelli博士的K23奖项的申请，他是一名血液学家，医学系终身教职流助理教授，匹兹堡大学成人镰状细胞性贫血项目主任。诺维利博士在良性血液学领域确立了自己的学术领袖地位，专注于镰状细胞疾病和止血方面的转化性研究。Novelli博士在血液学方面拥有丰富的研究经验，并因其在SCD方面的研究而成功获得研究奖项。他已经组建了一个由导师和顾问组成的多学科团队，以指导他的职业生涯走向独立，并协助完成本申请中提出的研究。这笔赠款将对实现以下职业目标至关重要：(1)发展实验室方法学方面的专门知识，对开展SCD的转化性研究至关重要；(2)进一步深入了解临床试验设计和人类疾病研究的基础生物统计学；以及(3)成为SCD血管生物学方面的专家。他计划通过完成匹兹堡大学赞助的临床研究理学硕士学位来补充他的研究活动，参加预定的关于拨款撰写和负责任的研究进行的培训活动，并参加他感兴趣的领域的国际会议。到这个K23奖项的资助期结束时，他将拥有几份第一次创作的同行评议出版物和大量的临界量数据，这些数据将构成RO1或其他同等赠款的核心。这一应用的重点是SCD的血管生物学。在SCD中，突变的血红蛋白S在脱氧时聚合，驱动红细胞依赖的血管闭塞和溶血。这些过程会导致血小板和止血活性、肺高压和血管疾病。仅表达人类和S珠蛋白的转基因基因敲除镰刀(BERK)小鼠在人类中模拟SCD，表现为NO生物利用度降低、NO介导的血管反应性受损和肺动脉高压。最近发现，血小板颗粒蛋白血栓反应蛋白-1(TSP1)通过与细胞表面受体CD47结合，有效地抑制生理性NO信号转导。Novelli博士的初步数据现在表明，SCD患者血浆中循环TSP1水平增加。支持这一建议的临床前研究也表明，TSP1-CD47配体受体的特异性相互作用不仅抑制内皮细胞中的NO信号转导，而且还增加了 活性氧(ROS)的产生和内皮素A(ETA)受体在平滑肌中的表达，都是导致肺动脉高压的典型的血管收缩和有丝分裂的病理信号通路。使用嵌合体交叉移植系统的大量初步数据证实，CD47在转基因镰状细胞小鼠的PH发生中起关键作用。这些发现为赠款的总体假设提供了信息，即在SCD患者的血小板激活过程中，循环血浆TSP1水平通过与TSP1结合而增加 CD47受体干扰肺血管内皮细胞NO的产生，刺激血管内皮细胞产生ROS和ET-1信号，从而导致SCD模型和人的肺动脉高压。这笔赠款还将探索这样一种假设，即治疗中断TSP1-CD47配体-受体相互作用将预防和逆转SCD的肺动脉高压。这一假说将通过以下目的得到验证：(1)首次检测SCD患者血浆中TSP1水平是否通过抑制NO信号、增加ROS和ETA水平而破坏血管内皮和血管收缩平衡；(2)利用独特的突变小鼠模型，确定循环TSP1和CD47在SCD相关血管病变和肺动脉高压中的作用，并测试TSP1-CD47阻断TSP1-CD47预防/缓解SCD相关性肺动脉高压的疗效；(3)检测SCD患者大队列中TSP1水平，探讨TSP1与血小板活化、肺动脉高压、血管疾病的关系。这一建议得到了用于研究假说的补充和丰富的工具集的支持，血管医学研究所内可用的大量资源， 匹兹堡大学致力于血管生物学的研究，并捕捉到了Novelli博士的导师在定义SCD病理生理学中的溶血(Mark Gladwin博士的实验室)和TSP1-CD47联系作为NO信号的上游调节因子(Jeff Isenberg博士的实验室)方面的开创性工作。因此，该通路的定义将为SCD的血管并发症提供一个新的治疗靶点。