Proteogenomics of dysregulated protein interaction networks in MTB infection

描述(申请人提供)：雾化结核分枝杆菌(MTB)很容易在人与人之间传播。分枝杆菌感染远端肺泡，面对先天反应，它们在那里不受约束地复制。随着适应性T细胞免疫的发展，结核分枝杆菌的生长得到了控制，但细菌并没有被根除。T细胞与结核杆菌感染的抗原提呈细胞(APC)之间的相互作用是机体逃避/抵抗宿主免疫以建立潜伏感染的核心。在这项应用中，针对RFA-HL-10-015“结核病潜伏和再激活机制的系统生物学方法”，我们汇集了蛋白质组学、计算机科学、生物信息学、遗传流行病学、免疫学、肺部生物学和肺部医学等领域的多学科专家团队，以获取一组独特的流行病学和临床特征良好的人群，这些人在美国、乌干达和南非具有结核杆菌暴露、感染和疾病的频谱，以便应用新的系统生物学方法来治疗结核杆菌潜伏感染。最近的研究表明，蛋白质组学方法旨在识别蛋白质-蛋白质相互作用网络，从而识别在疾病发病机制中起作用的功能亚网络。我们建议将这种方法应用于分析人类潜在的结核分枝杆菌感染(LTBI)，并将蛋白质组学结果与使用人类遗传和系统化学/细胞因子方法了解结核分枝杆菌发病机制的平行研究联系起来。这一建议的一般假设是，蛋白质组种子识别区分结核杆菌感染和疾病不同阶段的蛋白质的子网络，并提供对从下呼吸道感染到活动性结核病的机制的洞察(S)。其目的是：目的1：确定未感染结核分枝杆菌重度暴露家庭中单核巨噬细胞和CD4+T细胞中蛋白质-蛋白质相互作用亚网络失调的情况，并与未进展为疾病的LTBI患者和发展为结核病的患者进行比较。目的2：在与目标1相同的人群中，确定蛋白质-蛋白质相互作用亚网络与靶基因分析、外周血化学/细胞因子反应的关系。目的3：确定目标1和2中分析的蛋白质-蛋白质相互作用和化学/细胞因子亚网络中的哪一个与LTBI患者的肺CD4+T细胞和巨噬细胞最相关。这项建议结合了从美国、乌干达和南非的流行病学特征良好的结核杆菌感染者那里获得独特的临床标本(外周血细胞、血浆、支气管肺泡灌洗标本)，以及使用蛋白质组学、遗传流行病学和细胞因子生物学的专家，对下呼吸道感染及其发展为活动性结核病的多学科系统生物学方法。 公共卫生相关性：系统生物学非常适合于分析人类与结核分枝杆菌(MTB)之间的复杂相互作用，结核分枝杆菌是结核病(TB)的原因，当应用于结核杆菌暴露、感染和疾病的不同阶段的人的组织时，系统生物学尤其强大。使用系统生物学来寻找受结核杆菌暴露或感染干扰的关键宿主蛋白，使我们能够识别有可能发展为活动性结核病的人，并找出我们需要成为自然或通过接种疫苗预防结核病的人。

DESCRIPTION (provided by applicant): Aerosolized Mycobacterium tuberculosis (MTB) is readily transmitted from person to person. Mycobacteria infect the distal alveoli where they replicate unfettered in the face of innate responses. As adaptive T cell immunity develops, MTB growth is controlled but bacilli are not eradicated. The interactions between T cells and MTB infected antigen presenting cells (APC) are central for the organism to evade/resist host immunity to establish latent infection. In this application in response to RFA-HL-10-015 "Systems Biology Approach to the Mechanisms of TB Latency and Reactivation" we bring together a multidisciplinary team of experts in proteomics, computer science, bioinformatics, genetic epidemiology, epidemiology, immunology, lung biology and pulmonary medicine coupled to access to a unique set of cohorts of epidemiologically and clinically well characterized persons with the spectrum of MTB exposure, infection and disease in the US, Uganda and S. Africa, in order to apply novel systems biology approaches to latent MTB infection. Recent studies suggest that proteomic approaches aimed at identifying protein-protein interaction networks result in the identification of functional sub-networks with a role in disease pathogenesis. We propose to apply this approach to the analysis of latent MTB infection (LTBI) in humans, and to link proteomic results with parallel studies using human genetic and systemic chemo-/cytokine approaches to understanding MTB pathogenesis. The general hypothesis of this proposal is that proteomic seeds identify sub-networks of proteins that differentiate persons at different stages of MTB infection and disease, and provide insight into the mechanism(s) responsible for progression from LTBI to active TB. The aims are: Aim 1: To determine dysregulated protein-protein interaction sub-networks in mononuclear phagocytes and CD4+ T cells of persons in households heavily exposed to MTB who do not become infected compared to those with LTBI who do not progress to disease and those who develop TB. Aim 2: To determine the relationship between protein-protein interaction sub-networks to whole genome and targeted gene analyses, and peripheral chemo-/cytokine responses detected by multiplex chemo-/cytokine assays in the same population as in Aim 1. Aim 3: To determine which of the protein-protein interaction and chemo-/cytokine sub-networks analyzed in Aims 1 and 2 are most relevant for lung CD4+ T cells and macrophages from persons with LTBI. This proposal combines access to unique clinical specimens (peripheral blood cells, plasma, broncho- alveolar lavage specimens) from epidemiologically well characterized persons with MTB infection in US, Uganda and South Africa with experts in the use of proteomics, genetic epidemiology and cytokine biology for a multidisciplinary systems biology approach to LTBI and its progression to active TB. PUBLIC HEALTH RELEVANCE: Systems biology is ideally suited to analyze the complex interaction between humans and M. tuberculosis (MTB), the cause of tuberculosis (TB), and is particularly powerful when applied to tissues from persons in different stages of MTB exposure, infection and disease. Using systems biology to find key host proteins disturbed by MTB exposure or infection allows us to identify persons at risk for progression to active TB and find out what we need to be naturally or through vaccination protected against TB.