Immunoproteomic mechanisms of human macrophage resistance to Mycobacterium tuberculosis infection

人巨噬细胞抵抗结核分枝杆菌感染的免疫蛋白质组学机制

• 批准号: 10282139
• 负责人: Christine Anterasian
• 金额: $ 18.79万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-02 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10282139
• 关键词: AGFG1 gene; Address; Advisory Committees; Autophagocytosis; Award; Bacterial Proteins; Bioinformatics; Biological Assay; Cellular Immunology; Childhood; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Communicable Diseases; Data; Data Set; Development; Drug resistance in tuberculosis; Ensure; Exposure to; Family; Funding; Genes; Genetic; Genetic Polymorphism; Genetic Transcription; Goals; Growth; Guanosine Triphosphate Phosphohydrolases; Host Defense; Household; Human; Human Subject Research; Immune response; Individual; Infection; Interferons; International; Knowledge; Laboratories; Longitudinal Studies; Macrophage Activation; Mass Spectrum Analysis; Mediating; Membrane; Mentors; Mentorship; Mycobacterium tuberculosis; Natural Resistance; Outcome; Pathway Analysis; Pathway interactions; Patients; Pediatric Hospitals; Phenotype; Post-Translational Protein Processing; Process; Program Development; Proteins; Proteome; Proteomics; Pulmonary Tuberculosis; Quantitative Microscopy; Regulation; Research Personnel; Resistance; Role; Structure; Systems Biology; Techniques; Therapeutic; Training; Tuberculin Test; Tuberculosis; Ubiquitin; Ubiquitination; Uganda; Universities; Washington; Work; Writing; bioinformatics network; career development; cohort; cytokine; design; detection method; drug-sensitive; experimental study; improved; induced pluripotent stem cell; insight; large datasets; leukocyte activation; mRNA Expression; macrophage; meetings; novel; protein function; rab GTP-Binding Proteins; resistance mechanism; response; trafficking; transcriptomics

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.

项目总结/摘要 结核病（TB）是世界范围内的主要传染性杀手。接触结核分枝杆菌后 (Mtb)大多数人发展为无症状的潜伏性TB感染（LTBI表型）。然而，Anterasian博士 合作者发现，尽管家庭暴露于肺结核，但仍有7%的人没有 转换他们的结核菌素皮肤试验（TST）或干扰素-γ释放试验（IGRA），因此可以归类为 临床上对传统定义的LTBI具有抗性（“RSTR”表型）。揭示自然的机制 对Mtb感染的抗性可能提供独特的见解，可以告知宿主导向的 治疗（HDTs）。Anterasian博士分析了第一个全球结核病感染的原发性 人类巨噬细胞，并发现了46个差异丰富的蛋白质（DAP），定义了RSTR与LTBI 巨噬细胞对结核分枝杆菌的蛋白质组学反应通过将她的数据与合作者的数据相结合， 已经鉴定了可能经历Mtb诱导的翻译后修饰（PTM）和/或与Mtb相互作用的DAP。 结核杆菌蛋白质。她还使用生物信息学网络分析以及遗传和转录组学 来自同一患者队列的数据，以最终整理出19种蛋白质的列表，用于进一步的机制研究。在 特别地，GTP酶的Rab家族与Mtb细菌蛋白相互作用，经历Mtb诱导的泛素化， 并且是膜运输和自噬的关键调节剂，这是Mtb在 感染本提案的目的是确定巨噬细胞途径和蛋白质， 保护性RSTR反应。Anterasian博士假设RSTR个体促进巨噬细胞Mtb 通过DAP-Mtb蛋白相互作用和差异DAP调节的DAP介导途径的清除 泛素化在目标1中，Anterasian博士将研究Rab-dependent机制， 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 与结核分枝杆菌感染控制相关的蛋白质，它们的PTM，以及 阻力通过结合蛋白质组学，生物信息学和细胞免疫学方法，Anterasian博士 准备确定人类巨噬细胞对Mtb耐药性的关键途径，这些途径可以用HDTs靶向。 博士Anterasian是西雅图传染病部的儿科传染病研究员 儿童医院和华盛顿大学。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的指导培训; 2）正式 大数据集分析教学法（即蛋白质组学，统计遗传学，系统生物学）; 3） 设计蛋白质组学研究，人类受试者研究和科学写作; 4）结构化的机会， 向当地和国际科学界人士介绍她的工作;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 过渡到一位独立资助的结核病宿主应对专家，她将领导自己的实验室。