Immunoproteomic mechanisms of human macrophage resistance to Mycobacterium tuberculosis infection

项目摘要/摘要 结核病(TB)是全球头号传染病杀手。暴露于结核分枝杆菌后 (MTB)，大多数人发展为无症状的潜伏性结核病感染(LTBI表型)。然而，安特拉西亚博士的 合作者发现，大约7%的人，尽管家庭暴露于肺结核病，但没有 转换为结核菌素皮肤试验(TST)或干扰素γ释放试验(IGRA)，因此可分类为 临床上对传统定义的LTBI(“RSTR”表型)耐药。揭示了自然界的机制 对结核分枝杆菌感染的抵抗力可能提供独特的见解，可以为宿主定向感染的发展提供信息。 治疗公司(HDTS)。Anterasian博士分析了全球首个结核分枝杆菌感染原发病例的蛋白质组数据集 人巨噬细胞发现了46个差异丰富的蛋白质(DAP)，它们定义了RSTR与LTBI 巨噬细胞对结核分枝杆菌的蛋白质组反应。通过将她的数据与她的合作者的数据相结合，Anterasian博士 已确定可能经历Mtb诱导的翻译后修饰(PTM)和/或与 结核分枝杆菌细菌蛋白。她还使用了生物信息网络分析以及基因和转录组分析 来自同一患者队列的数据，以最终整理出19种蛋白质的清单，用于进一步的机制研究。在……里面 特别是，Rab家族的GTP酶与Mtb细菌蛋白相互作用，经历Mtb诱导的泛素化， 和是膜运输和自噬的关键调节者，这是结核分枝杆菌在 感染。这项建议的目标是定义巨噬细胞途径和蛋白质，以表征 保护性RSTR反应。Anterasian博士假设RSTR个体促进巨噬细胞Mtb DAP-Mtb蛋白相互作用和差异DAP调节的DAP介导的清除 泛素化。在目标1中，Anterasian博士将研究针对结核分枝杆菌的依赖于Rab的保护机制 巨噬细胞中的Mtb蛋白以及Mtb蛋白如何破坏Rab功能。在目标2中，Anterasian博士将确定候选人 与控制结核分枝杆菌感染相关的蛋白质、它们的PTM以及泛素依赖的机制 抵抗。通过结合蛋白质组学、生物信息学和细胞免疫学的方法，Anterasian博士做得很好 准备确定人类巨噬细胞对结核分枝杆菌耐药的关键途径，这些途径可以作为HDTS的靶点。 Anterasian博士是西雅图传染病科的儿科传染病研究员 儿童医院和华盛顿大学。此外，她还提出了一个全面的职业生涯 发展计划包括：1)蛋白质组学和细胞免疫学方面的指导培训；2)正式的 大数据集分析(即蛋白质组学、统计遗传学、系统生物学)的教学；3)在 设计蛋白质组学研究、人类受试者研究和科学写作；4)有组织的机会 向当地和国际科学受众介绍她的工作；以及5)科学咨询委员会会议 确保科学和职业发展进步。在这个奖项结束时，Anterasian博士将 过渡到MTB东道主响应中的一名独立资助的专家，该专家将指导自己的实验室。

PROJECT SUMMARY / ABSTRACT Tuberculosis (TB) is the leading infectious killer worldwide. Upon exposure to Mycobacterium tuberculosis (Mtb), most people develop asymptomatic latent TB infection (LTBI phenotype). However, Dr. Anterasian's collaborators identified about 7% of individuals who, despite household exposure to pulmonary TB, do not convert their tuberculin skin test (TST) or interferon-γ release assay (IGRA), and thus can be classified as clinically resistant to traditionally defined LTBI (“RSTR” phenotype). Uncovering mechanisms of natural resistance to Mtb infection may provide unique insights that can inform the development of host-directed therapeutics (HDTs). Dr. Anterasian has analyzed the first global proteomic dataset of Mtb-infected primary human macrophages and discovered 46 differentially abundant proteins (DAPs) that define the RSTR vs LTBI macrophage proteomic response to Mtb. By integrating her data with those of her collaborators, Dr. Anterasian has identified DAPs that may undergo Mtb-induced post-translational modifications (PTMs) and/or interact with Mtb bacterial proteins. She has also used bioinformatic network analyses as well as genetic and transcriptomic data from the same patient cohort to ultimately curate a list of 19 proteins for further mechanistic studies. In particular, the Rab family of GTPases interact with Mtb bacterial proteins, undergo Mtb-induced ubiquitination, and are key regulators of membrane trafficking and autophagy, which are pathways Mtb exploits during infection. The objective of this proposal is to define macrophage pathways and proteins that characterize the protective RSTR response. Dr. Anterasian hypothesizes that RSTR individuals promote macrophage Mtb clearance through DAP-mediated pathways modulated by DAP-Mtb protein interactions and differential DAP ubiquitination. In Aim 1, Dr. Anterasian will investigate Rab-dependent mechanisms of protection against Mtb in macrophages and how Mtb proteins subvert Rab function. In Aim 2, Dr. Anterasian will identify candidate proteins associated with control of Mtb infection, their PTMs, and ubiquitin-dependent mechanisms of resistance. By combining proteomic, bioinformatic, and cellular immunology approaches, Dr. Anterasian is well poised to identify key pathways in human macrophage resistance to Mtb that can be targeted with HDTs. Dr. Anterasian is a Pediatric Infectious Disease Fellow in the Division of Infectious Diseases at Seattle Children's Hospital and the University of Washington. She additionally proposes a comprehensive career development program that includes: 1) mentored training in proteomics and cellular immunology; 2) formal didactics in large data set analyses (i.e. proteomics, statistical genetics, systems biology); 3) mentorship in the design of proteomic studies, human subjects research, and scientific writing; 4) structured opportunities to present her work to local and international scientific audiences; and 5) Scientific Advisory Committee meetings that ensure scientific and career development progress. By the conclusion of this award, Dr. Anterasian will transition to an independently-funded expert in the Mtb host response who will direct her own laboratory.