NLRP3 Inflammasome Activation and Mitochondrial Function in the setting of Aging and HIV Infection

项目摘要/摘要： 老龄化和老年艾滋病毒感染人群的特征都是代谢率增加。 综合征(定义为腹型肥胖、血脂异常、胰岛素抵抗和高血压)。值得注意的是， 代谢综合征与失调的、与年龄相关的促炎环境有关- 称为“炎性老化”--以细胞因子、急性期反应物和凝血水平升高为特征。 各种因素。两种病原体相关分子模式对先天免疫受体的慢性刺激 (PAMPS)和损伤相关的分子模式(DAMP)被认为是与年龄相关的 慢性炎症，但代谢综合征的发病机制 老龄化和艾滋病毒疾病的研究仍然是该领域尚未完全了解的知识鸿沟。NLRP3(点头- 与受体吡咯结构域一样，炎症体是一种细胞内蛋白质复合体，是 并介导依赖于caspase-1的前IL-1b和前IL-18对其 激活的表单。虽然NLRP3炎症体被PAMPs激活，但有越来越多的证据表明， NLRP3通过DAMPS作为宿主新陈代谢的感受器的作用，通过广泛的 代谢物。此外，NLRP3炎症体的激活依赖于线粒体的功能。NLRP3 炎症小体与胰岛素抵抗和其他代谢综合征的发展有关 小鼠模型，并在老年人和感染艾滋病毒的成年人中进行了最低限度的探索。目的 本研究旨在确定年龄和HIV感染对NLRP3炎症体的影响，并探讨其作用机制。 与线粒体功能的关系通过比较以下受试组，年轻人(21-35岁)， 以及有和没有艾滋病毒感染的老年人(≥60岁)。目标1试图描述NLRP3的特征 外周血髓系细胞炎症小体及其与线粒体功能的关系 脂肪组织。目标2试图描述由激活蛋白2诱导的代谢途径 外周血髓系细胞中NLRP3炎症体的RNA测序和细胞周期分析 脂肪组织。来自这两个目标的数据将结合临床特征进行收集，包括 代谢综合征的组成部分。我们的假设是，年龄的增长和艾滋病毒的感染将导致 NLRP3炎症体功能失调与激活有关 线粒体功能障碍-最终导致老年人和老年人代谢综合征的发展 感染艾滋病毒的成年人。候选人萨帕塔博士是耶鲁大学医学院传染病内科医生。 医学，谁已经成立了一个跨学科的指导委员会，在免疫学，老龄化， 和新陈代谢。该培训方案与包括导师制在内的职业发展计划相结合。 和免疫新陈代谢的教学培训，另一个重点是学习测序分析 数据，从而提供了使PI能够申请R01奖的工具。

Project Summary/Abstract: Both the aging and the aging HIV-infected population are characterized by increased rates of metabolic syndrome (defined by abdominal obesity, dyslipidemia, insulin resistance and hypertension). Notably, metabolic syndrome is associated with the dysregulated, age-associated pro-inflammatory environment— termed “Inflamm-aging”—characterized by elevated levels of cytokines, acute phase reactants, and clotting factors. Chronic stimulation of innate immune receptors by both pathogen-associated molecular patterns (PAMPs) and damage associated molecular patterns (DAMPs) is thought to contribute to age-associated chronic inflammation, but the mechanisms underlying the pathogenesis of metabolic syndrome in the context of aging and HIV disease remain an incompletely understood knowledge gap in the field. The NLRP3 (NOD- like receptor pyrin domain-containing 3) inflammasome is an intracellular protein complex, that is part of the innate immune response and mediates the caspase-1-dependent cleavage of pro-IL-1b and pro-IL-18 to their activated forms. While the NLRP3 inflammasome is activated by PAMPs, there is increasing evidence for a role of NLRP3 as a sensor of host metabolism via DAMPs, as shown by NLRP3 activation by a wide range of metabolites. Moreover, NLRP3 inflammasome activation is dependent on mitochondrial function. The NLRP3 inflammasome has been linked to the development of insulin resistance and other metabolic syndromes in mouse models, and has been minimally explored in both older adults and HIV-infected adults. The purpose of this proposal is to determine the effects of age and HIV infection on the NLRP3 inflammasome, and its relationship with mitochondrial function by comparing the following groups of subjects, young adults (21-35), and older adults (≥ 60 yrs) with and without HIV-infection. Aim 1 seeks to characterize the NLRP3 inflammasome and its relationship with mitochondrial function, in myeloid cells from peripheral blood and adipose tissue. Aim 2 seeks to characterize the metabolic pathways that are induced with activation of the NLRP3 inflammasome through RNA sequencing and CyTOF in myeloid cells from peripheral blood and adipose tissue. Data from both aims will be collected in conjunction with clinical characteristics including the components of metabolic syndrome. Our hypothesis is that increased age and HIV infection will result in dysregulated NLRP3 inflammasome function at baseline and with activation that is linked to mitochondrial dysfunction—ultimately contributing to the development of metabolic syndrome in older and HIV-infected adults. The candidate, Dr. Zapata is an Infectious Disease physician at the Yale school of medicine, who has put together an interdisciplinary mentorship committee with expertise in immunology, aging, and metabolism. This training proposal is coupled with a career development plan that includes mentorship and didactic training in Immuno-metabolism, with an additional focus on learning the analysis of sequencing data, thus providing the tools that will allow the PI to apply for an R01 award.