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: 9094734
• 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/9094734
• 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; NADP; NADPH Oxidase; Null Lymphocytes; Oils; Oxidases; Paper; 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; Smooth Muscle Myocytes; Source; Staining method; Stains; Subendothelial Layer; Superoxides; Technology; Testing; Therapeutic; Thrombosis; Thrombospondin 1; Tissues; Training; Transplantation; United States; United States National Institutes of Health; Vascular Smooth Muscle; Woman; Writing; atherogenesis; career; cost; cytochrome c; field study; gel electrophoresis; health care delivery; improved; in vivo; insight; macrophage; men; multidisciplinary; novel; oxidized low density lipoprotein; prevent; programs; receptor; research study; response; skills; skills training; therapeutic target

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 HL114648-01 的重新提交。 Csanyi 博士正在将自己定位为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 (TSP1) 介导的 CD47 信号传导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) 合并截然不同的 NADPH 氧化酶 (Nox) 和 TSP1 领域，目标是开辟新的发现途径并建立 Csanyi 博士的独立研究计划； 3) 成为心血管和 TSP1 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。随着 Csanyi 博士走向科学独立，他将从参加以下研讨会中受益匪浅：科学管理和领导力课程、管理职业方向和资助写作研讨会。到本次 K99/R00 奖项资助期结束时，预计 Csanyi 博士将发表多篇高影响力的第一作者和最后作者论文，并成功竞争后续 NIH 资助。该研究计划的总体目标是研究一条新的机制途径，将巨噬细胞中病理性 TSP1 介导的 CD47 激活与 Nox 激活和泡沫细胞形成联系起来，从而促进动脉粥样硬化斑块的演变、斑块不稳定和血栓形成。该资助的总体假设是，TSP1 激活 CD47 会促进巨噬细胞中 Nox 衍生的活性氧 (ROS) 的产生增加，从而增加 LDL 的氧化及其吞噬作用，从而导致泡沫细胞形成（目标 1）并引发动脉粥样硬化（目标 2）。还提出巨噬细胞中 CD47 驱动的 ROS 会增加基质金属蛋白酶活性，导致斑块不稳定并最终导致动脉粥样硬化斑块破裂和血栓形成（目标 3）。将使用细胞色素 c、电子顺磁共振和 2-羟基乙锭定量来测量 ROS。将使用 CD47 结合序列、CD47 无效巨噬细胞和 CD47 基因沉默吗啉来研究 CD47 的作用。 Nox 无效细胞和 siRNA 技术将用于表征 TSP1 刺激的 ROS 来源。将创建独特的 CD47 null ApoE null 骨髓嵌合体，以研究巨噬细胞 CD47 对斑块破裂和血栓形成的贡献。将评估 CD47 吗啉减弱斑块进展和防止破裂的治疗潜力。这项努力非常重要，因为它有潜力 a) 开辟一个全新的研究领域； b) 提供新证据表明 TSP1-CD47 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剂； c) 将 TSP1-CD47-Nox 连接确定为动脉粥样硬化和其他心血管疾病的潜在治疗靶点。