A Novel Role of Macrophage TSP1-CD47 Signaling in Atherosclerosis

巨噬细胞 TSP1-CD47 信号传导在动脉粥样硬化中的新作用

• 批准号: 9294101
• 负责人: Gabor Csanyi
• 金额: $ 24.9万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9294101
• 关键词: Academic Training; Antibodies; Aorta; Apolipoprotein E; Applications Grants; Arterial Fatty Streak; Atherosclerosis; Attenuated; Award; Binding; Biology; Blood Vessels; Bone Marrow; CD47 gene; Cardiovascular Diseases; Cardiovascular system; Cells; Cessation of life; Chimera organism; Coronary Arteriosclerosis; Coronary Thrombosis; Coronary artery; Coronary heart disease; Deposition; Development; Disease; Educational workshop; Electron Spin Resonance Spectroscopy; Enzyme-Linked Immunosorbent Assay; Event; Evolution; Foam Cells; Free Radicals; Functional disorder; Funding; Gap Junctions; Gene Silencing; Generations; Goals; Grant; Human; In Vitro; Infiltration; Inflammatory; Ischemia; Knowledge; Leadership; Lesion; Link; Lipids; Low Density Lipoprotein oxidation; Matrix Metalloproteinases; Measures; Mediating; Mentors; Molecular; Mus; Muscle Cells; NADP; NADPH Oxidase; Null Lymphocytes; Oils; Oxidases; Oxides; Paper; Pathologic; Pathway interactions; Phagocytosis; Phase; Play; Positioning Attribute; Process; Production; Proteins; Publishing; Reactive Oxygen Species; Reporting; Research; Research Personnel; Research Project Grants; Role; Rupture; Science; Scientist; Secure; Seminal; Shoulder; Signal Transduction; Small Interfering RNA; Source; Staining method; Stains; Subendothelial Layer; Superoxides; Technology; Testing; Therapeutic; Thinness; Thrombosis; Thrombospondin 1; Tissues; Training; Transplantation; United States; United States National Institutes of Health; Vascular Smooth Muscle; Woman; Writing; atherogenesis; career; cost; cytochrome c; experimental study; field study; gel electrophoresis; health care delivery; improved; in vivo; inflammatory milieu; insight; macrophage; men; multidisciplinary; novel; oxidation; oxidized low density lipoprotein; prevent; programs; receptor; response; skills; skills training; therapeutic target; thrombospondin 2

DESCRIPTION (provided by applicant): This is a resubmission of K99 HL114648-01. Dr. Csanyi is establishing himself as an independent investigator in the field of cardiovascular science with specific focus on the role of thrombospondin-1 (TSP1)-mediated CD47 signaling in atherosclerosis and vascular free radical biology. This grant will be critical to achieve the following short- and long-term objectives: 1) to acquire additional scientific training in skills ad knowledge; 2) to merge the yet distinct NADPH oxidase (Nox) and TSP1 fields with the goals toward opening up new avenues of discovery and establishing Dr. Csanyi's independent research program; and 3) to become an independently funded scientist at the forefront of cardiovascular and TSP1-related research. Dr. Csanyi has assembled a multidisciplinary team, including his mentor, collaborators, and advisors to guide his career towards independence and assist with the completion of the research proposed in this application. As Dr. Csanyi moves towards his scientific independence he will greatly benefit from attending the following seminars: Course in Scientific Management and Leadership, Managing the Direction of Your Career, and Grant Writing workshop. By the end of the funding period of this K99/R00 award, it is expected that Dr. Csanyi will have published several high-impact first and last author papers and successfully competed for subsequent NIH funding. The overall objective of the research plan is to investigate a new mechanistic pathway linking pathological TSP1-mediated CD47 activation in macrophages with Nox activation and foam cell formation, contributing to the evolution of atherosclerotic plaques, plaque destabilization and thrombosis. The grant's overarching hypothesis is that CD47 activation by TSP1 promotes increased Nox-derived reactive oxygen species (ROS) production in macrophages, increasing oxidation of LDL and its phagocytosis, resulting in foam cell formation (Aim 1) and initiation of atherosclerosis (Aim 2). It is also proposed that CD47-driven ROS in macrophages increases matrix metalloproteinase activity leading to plaque destabilization and ultimate rupture of atherosclerotic plaques and thrombosis (Aim 3). ROS will be measured using cytochrome c, electron paramagnetic resonance, and quantification of 2-hydroxyethidium. The role of CD47 will be investigated using CD47 binding sequences, CD47 null macrophages, and CD47 gene silencing morpholinos. Nox null cells and siRNA technology will be used to characterize the source of TSP1-stimulated ROS. Unique CD47 null  ApoE null bone marrow chimeras will be created to investigate the contribution of macrophage CD47 to plaque rupture and thrombosis. The therapeutic potential of CD47 morpholinos to attenuate plaque progression and prevent rupture will be evaluated. This endeavor is highly significant because it has the potential to a) open up an entirely new field of study; b) provide new evidence that TSP1-CD47 signaling is a stimulator of macrophage ROS, foam cell formation and unstable plaque; and c) identify the TSP1-CD47-Nox nexus as a potential therapeutic target in atherosclerosis and other cardiovascular disorders.

描述（由申请人提供）：这是K99 HL 114648 -01的重新提交。Csanyi博士是心血管科学领域的独立研究者，特别关注血小板反应蛋白-1（TSP 1）介导的CD 47信号在动脉粥样硬化和血管自由基生物学中的作用。这笔赠款对于实现以下短期和长期目标至关重要：1）获得技能和知识方面的额外科学培训; 2）合并尚未区分的NADPH氧化酶（Nox）和TSP 1领域，以开辟新的发现途径并建立Csanyi博士的独立研究计划;和3）成为心血管和TSP 1相关研究前沿的独立资助科学家。Csanyi博士组建了一个多学科团队，包括他的导师，合作者和顾问，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。Csanyi走向他的科学独立性，他将大大受益于参加以下研讨会：科学管理和领导力课程，管理你的职业生涯的方向，并授予写作研讨会。到K99/R 00奖的资助期结束时，预计Csanyi博士将发表多篇具有高影响力的第一作者和最后作者论文，并成功竞争随后的NIH资助。该研究计划的总体目标是研究一种新的机制途径，将巨噬细胞中病理性TSP 1介导的CD 47活化与Nox活化和泡沫细胞形成联系起来，从而促进动脉粥样硬化斑块、斑块不稳定和血栓形成的演变。格兰特的首要假设是，由TSP 1激活的CD 47促进巨噬细胞中NOx衍生的活性氧（ROS）产生增加，增加LDL的氧化及其吞噬作用，导致泡沫细胞形成（Aim 1）和动脉粥样硬化的起始（Aim 2）。还提出巨噬细胞中CD 47驱动的ROS增加基质金属蛋白酶活性，导致斑块不稳定和动脉粥样硬化斑块的最终破裂和血栓形成（目的3）。将使用细胞色素c、电子顺磁共振和2-羟基噻啶定量测定ROS。将使用CD 47结合序列、CD 47无效巨噬细胞和CD 47基因沉默吗啉代研究CD 47的作用。Nox空细胞和siRNA技术将用于表征TSP 1刺激的ROS的来源。唯一CD 47空  将创建ApoE无效骨髓嵌合体以研究巨噬细胞CD 47对斑块破裂和血栓形成的贡献。将评估CD 47吗啉代减弱斑块进展和预防破裂的治疗潜力。这一奋进是非常重要的，因为它有可能a）开辟一个全新的研究领域; B）提供新的证据，证明TSP 1-CD 47信号传导是巨噬细胞ROS、泡沫细胞形成和不稳定斑块的刺激物;和c）鉴定TSP 1-CD 47-Nox连接作为动脉粥样硬化和其他心血管疾病的潜在治疗靶点。