The Contribution of T cells to the Pathogenesis of Atherosclerosis in Older Adults

项目总结/摘要 动脉粥样硬化是一种慢性炎症性疾病，影响约70%的老年男性和女性 ≥ 65岁，引起心脏病发作、中风、心力衰竭、肢体丧失、痴呆，甚至死亡。老年人 由于免疫衰老，可能更容易发生动脉粥样硬化及其不良后遗症， 随着免疫系统的逐渐恶化，由于自然老化。虽然许多研究表明， 血液中检测到的促炎性T细胞增加了斑块进展和破裂的风险， 已知动脉粥样硬化中抗原特异性T细胞活化和/或T细胞免疫衰老 斑块改变了T细胞亚群的功能，并促进了更脆弱的表型。迫切需要 为了更好地理解T细胞衰老导致的发病机制， 动脉粥样硬化;没有这些信息，药物治疗旨在减轻衰老免疫的影响， 系统对动脉粥样硬化及其不良后果的研究将是无法实现的。由于发展单一 细胞技术和下一代测序，专门设计用于更好地了解T细胞生物学 由申请人的导师，T细胞克隆性和衰老是否有助于 老年人动脉粥样硬化的发展及其不良后遗症现在可以得到解答。来 在过去的一年里，申请人已经应用这些技术来开始回答这个问题，并发现 四名老年人动脉粥样硬化斑块中克隆性免疫衰老T细胞的存在。以下 计划将解决这一问题：目标1：导致动脉粥样硬化的克隆性T细胞群， 老年人在这个目标中，将对单细胞水平的T细胞受体（TCR）进行测序和表型分析 采用由申请人的导师开创并已被申请人成功实施的方法; 目的2：鉴定激活动脉粥样硬化斑块中寡克隆T细胞扩增的抗原 老年动脉粥样硬化患者。目标1中生成的TCR数据将被组织， 通过申请人导师开发的经过充分验证的算法进行抗原鉴定分析;以及Aim 3：表征区分不稳定与稳定疾病的抗原特异性T细胞。抗原特异性t 细胞将使用由申请人开发的新型肽-MHC十二聚体技术来表征。 导师该提议是创新的，因为它通过利用单细胞技术的进步而脱离现状。 测序，计算分析和十二聚体技术，使免疫基础的发展 直接针对斑块进展和破裂的治疗，开辟了新的研究视野。拟议 研究是重要的，因为它有望纵向推进和扩大我们对如何 抗原特异性T细胞活化和/或免疫衰老改变T细胞功能并促进斑块形成 进展和破裂。这些知识将使基于免疫的疗法的发展能够靶向 老年人的动脉粥样硬化

PROJECT SUMMARY/ABSTRACT Atherosclerosis is a chronic inflammatory disorder that affects approximately 70% of older men and women ≥65 years old, causing heart attack, stroke, heart failure, limb loss, dementia, and even death. Older adults may be more susceptible to atherosclerosis and its adverse sequelae because of immune senescence, defined as the gradual deterioration of the immune system due to natural aging. While many studies have shown that pro-inflammatory T cells detected in the blood increases the risk of plaque progression and rupture, little is known about how antigen specific T cell activation and/or T cell immune senescence in the atherosclerotic plaque alters the function of T cell subsets and promotes a more vulnerable phenotype. There is a critical need to better understand the mechanisms by which T cell senescence contributes to the pathogenesis of atherosclerosis; without this information, pharmacotherapy designed to mitigate the effects of an aging immune system on atherosclerosis and its adverse sequelae will be unattainable. Because of the development of single cell technology and next generation sequencing that is specifically designed to better understand T cell biology by the applicant's mentor, the fundamental question of whether T cell clonality and senescence contributes to the development of atherosclerosis and its adverse sequelae in older adults can now be answered. Over the past year, the applicant has applied these techniques to begin answering this question and has found the presence of clonal, immunosenescent T cells in the atherosclerotic plaque in four older adults. The following plan will address this problem: Aim 1: Validate clonal T cell populations that cause atherosclerosis in older adults. In this aim, T cell receptors (TCRs) at the single cell level will be sequenced and phenotyped using an approach pioneered by the applicant's mentor and already successfully performed by the applicant; Aim 2: Identify antigen(s) that activate oligoclonal T cell expansion in atherosclerotic plaque isolated from older patients with advanced atherosclerosis. TCR data generated in Aim 1 will be organized and analyzed for antigen identification by a well-validated algorithm developed by the applicant's mentor; and Aim 3: Characterize antigen-specific T cells that distinguish unstable from stable disease. Antigen specific T cells will be characterized using novel peptide-MHC dodecamer technology developed by the applicant's mentor. This proposal is innovative because it departs from the status quo by utilizing advances in single cell sequencing, computational analysis, and dodecamer technology to enable the development of immune based therapy that directly targets plaque progression and rupture, opening new research horizons. The proposed research is significant, because it is expected to vertically advance and expand our understanding of how antigen specific T cell activation and/or immune senescence alters T cell function and promotes plaque progression and rupture. Such knowledge will enable the development of immune-based therapies to target atherosclerosis in older adults.