A Mechanistic and Translational Research Program Linking Impaired Resolution, Defective Efferocytosis, and Clonal Hematopoiesis to the Formation of Clinically Dangerous Atherosclerotic Plaques

R35提案是一项全面的长期计划，旨在探索新的机制， 与引起急性动脉粥样硬化的独特类型的动脉粥样硬化斑块的形成有关的治疗概念 心血管疾病（CVD）。对青年科学家的培训和指导也是该计划的一个关键部分。 PI多年来一直不间断地获得多项NIH赠款，发表关于动脉粥样硬化的文章， 心脏代谢疾病的影响力最高的期刊，并有一个非常成功的记录，培训年轻 科学家是独立的学术研究者。该计划将探索新的，高度相关的概念 与存在主要差距的四个关键领域有关：（i）炎症消退和红细胞增多症（ 死细胞）;（ii）临床上最重要的少数动脉粥样硬化病变的病理生理学;（iii） M细胞中的氨基酸代谢，因为它与高负荷的红细胞增多症有关;和（iv）衰老相关的克隆 造血（CH）。损害炎症消退的过程，与促进炎症消退的过程不同 炎症本身和缺陷性红细胞增多症促进临床相关的坏死性薄帽细胞的形成， 斑块该实验室的新工作表明，（i）M细胞内化多个凋亡细胞（高负荷）的能力， （ii）与巨噬细胞代谢相关的途径， 凋亡细胞衍生的氨基酸对于高负荷的细胞凋亡是关键的。另一个令人兴奋的新概念 初步数据支持的是，受损的红细胞增多症和消退会因CH而加重， 成为动脉粥样硬化CVD的主要年龄相关危险因素。该计划的总体愿景是研究 这些新的领域，首先使用（i）小鼠和人类的M β细胞，以阐明深入的机制;和（ii）遗传 改变小鼠以测试晚期斑块进展中的因果关系。R35还将探索治疗潜力， 这些想法在临床前模型，与假设，解决调解人治疗将有疗效， 安全性优于常规抗炎治疗。PI将使用灵活性和连续性 R35计划的目标是进入与人类研究相关的新方向。通过丰富的合作者网络 在哥伦比亚大学和其他地方，包括哥伦比亚大学的心血管和代谢精准医学项目， 该计划将把新发现应用于（i）人类动脉粥样硬化的分析;（ii）人类遗传学，包括 通过与PROMIS研究合作，在分解/胞吐途径中存在基因科斯的受试者; 和（iii）用人单核细胞和iPSC衍生的MIBs进行的研究，所述MIBs适合CRISPR/Cas9介导的 基因工程，由该计划的机械和人类基因数据的指导。通过灵活性 R35计划的前瞻性和前瞻性，所提出的机械工作和人类工作的结合 研究将提供一种力量组合，以填补危险斑块形成的关键空白;设计新的 治疗策略;并在这一关键研究领域培训和指导年轻科学家。 1

This R35 proposal represents a comprehensive, long-term program that explores new mechanisms and therapeutic concepts related to the formation of the unique types of atherosclerotic plaques that cause acute cardiovascular disease (CVD). The training and mentoring of young scientists is also a key part of this program. The PI has held multiple NIH grants without interruption for many years, publishes on atherosclerosis and cardiometabolic disease in the highest impact journals, and has a highly successful record of training young scientists to be independent academic researchers. The program will explore new, highly interrelated concepts related to four key areas in which major gaps exist: (i) inflammation resolution and efferocytosis (clearance of dead cells); (ii) pathophysiology of the minority of atherosclerotic lesions that are most clinically important; (iii) amino acid metabolism in Ms as it relates to high-burden efferocytosis; and (iv) aging-related clonal hematopoiesis (CH). Processes that impair inflammation resolution, which are distinct from those that promote inflammation per se, and defective efferocytosis promote the formation of clinically relevant necrotic, thin-capped plaques. The lab's new work indicates that (i) the ability of Ms to internalize multiple apoptotic cells (high-burden efferocytosis) is critical to avoid necrotic plaques (Cell 2017); and (ii) a pathway related to M metabolism of apoptotic cell-derived amino acids is critical for high-burden efferocytosis. Another exciting new concept supported by preliminary data is that impaired efferocytosis and resolution are exacerbated by CH, which is emerging as a major age-related risk factor for atherosclerotic CVD. The overall vision of the program is to study these new areas by first using (i) mouse and human Ms to elucidate in-depth mechanisms; and (ii) genetically altered mice to test causation in advanced plaque progression. The R35 will also explore the therapeutic potential of these ideas in pre-clinical models, with the hypothesis that resolution mediator therapy will have efficacy and safety advantages over conventional anti-inflammatory therapy. The PI will then use the flexibility and continuity of the R35 program to move into new directions related to human studies. Through a rich network of collaborators at Columbia and elsewhere—including Columbia's Cardiovascular and Metabolic Precision Medicine program— the program will apply the new discoveries to (i) analyses of human atheroma; (ii) human genetics, including subjects with KOs of genes in resolution/efferocytosis pathways through a collaboration with the PROMIS study; and (iii) studies with human monocyte- and iPSC-derived Ms that are amenable to CRISPR/Cas9-mediated genetic engineering, as guided by the program's mechanistic and human genetic data. Through the flexibility and forward-looking nature of the R35 program, the combination of the proposed mechanistic work and human studies will provide a power combination to fill in critical gaps in how dangerous plaque form; to devise novel therapeutic strategies; and to train and mentor young scientists in this critical area of research. 1