Human macrophage variation & TB pathogenesis

结核分枝杆菌暴露后的临床结果的变化范围从耐药性感染播散性疾病。 人类遗传和细胞机制的耐药性和传播在很大程度上是未知的， 提供了对新的治疗策略的深入了解。在越南，我们招募并检查了大量的PTB患者， 对宿主和病原体进行详细的免疫遗传学研究， 疾病的发展。在乌干达过去20年的一项大型结核病家庭接触研究中，我们发现， 约9%的密切成年家庭接触者持续TST和干扰素释放试验（IGRA） 在长期随访期间呈阴性，似乎对传统定义的LTBI（RSTR）有抵抗力。与 通过对结核分枝杆菌感染的单核细胞的转录和蛋白质组分析，我们发现了与结核分枝杆菌感染相关的基因和途径。 与LTBI相比，RSTR富集。一些基因在两个数据集中差异富集，包括 RAB 11B，一个参与囊泡运输的基因，在RSTR巨噬细胞中增加， 在人类-M。结核蛋白相互作用（PPI）网络。此外，通过细胞GWAS 在结核分枝杆菌感染的巨噬细胞中，我们发现了与IL-1β相关的人类多态性， 在以前不知道调节IL-1 β的基因中的表达。这些发现表明新的基因和 这些变体全面调节人Mtb诱导的IL-1β，IL-1β是一种促进Mtb在 巨噬细胞和鼠体内模型。在RSTR和LTBI（乌干达）的遗传关联研究中， TBM & PTB（越南），我们在几个候选基因中发现了多态性（与RSTR或TB相关 疾病或巨噬细胞IL-1 β调节）与临床结果密切相关。所有这些 数据支持我们的假设，即结核分枝杆菌诱导的巨噬细胞反应是遗传调节的， 不同的临床结果。然而，我们的知识有许多空白。首先，结核病引起的后， 来自RSTR、LTBI和TB疾病个体的巨噬细胞的翻译修饰（PTM）谱是 未知第二，全球人类基因调节剂（基因和变体）的结核分枝杆菌诱导的巨噬细胞抗- 微生物途径大多未知。第三，人类结核分枝杆菌耐药的分子和细胞机制 和传播几乎完全未知，包括Mtb菌株变异在发病机制中的作用。 为了解决这些差距，我们将使用基因组学，遗传学和蛋白质组学方法来分析人类巨噬细胞 并发现与临床结果相关和/或与Mtb菌株相关的差异丰度PTM， 依赖。然后，我们将使用遗传和细胞策略来发现抗微生物药物的全球调节剂。 巨噬细胞对Mtb感染的反应，并研究这些基因及其变体如何调节巨噬细胞 在结核分枝杆菌菌株变异的背景下发挥作用。我们的主要目标是发现Mtb之间的新漏洞 和巨噬细胞，从而告知疾病异质性的机制， 临床管理和有效的宿主导向疗法和疫苗的开发。

Variation in clinical outcomes after Mtb exposure ranges from resistance to infection to disseminated disease. Human genetic and cellular mechanisms of resistance and dissemination are largely unknown and may provide insight into novel therapeutic strategies. In Vietnam, we enrolled and examined large cohorts of PTB and TBM subjects with detailed immunogenetic studies of both the host and pathogen to discover determinants of disease progression. In Uganda in a large TB household contact study over the past 20 years, we found that ~9% of close adult household contacts remained persistently TST and Interferon- Release Assay (IGRA) negative during extended follow-up and appear to be resistant to traditionally defined LTBI (RSTR). With transcriptional and proteomic profiling of Mtb-infected monocytes, we discovered genes and pathways that are enriched in RSTR compared to LTBI. Some genes were differentially enriched in both datasets, including RAB11B, a gene involved in vesicle trafficking, which was increased in RSTR macrophages and also identified in the human-M. tuberculosis protein-protein interaction (PPI) network. Furthermore, with a cellular GWAS approach in Mtb-infected macrophages, we discovered human polymorphisms associated with IL-1β expression that are in genes not previously known to regulate IL-1. These findings suggest new genes and variants that globally regulate human Mtb induced IL-1β, a key cytokine that promotes control of Mtb in macrophages and in murine in vivo models. In genetic association studies in RSTRs & LTBI (Uganda) and TBM & PTB (Vietnam), we found polymorphisms in several candidate genes (associated with RSTR or TB disease or macrophage IL-1 regulation) that were strongly associated with clinical outcomes. Together, these data support our hypothesis that Mtb-induced macrophage responses are genetically regulated and associated with different clinical outcomes. However, there are many gaps in our knowledge. First, the Mtb-induced post- translational modification (PTM) profiles of macrophages from RSTR, LTBI, and TB disease individuals are unknown. Second, the global human genetic regulators (genes and variants) of Mtb-induced macrophage anti- microbial pathways are mostly unknown. Third, molecular and cellular mechanisms of human Mtb resistance and dissemination are almost completely unknown, including the role of Mtb strain variation in pathogenesis. To address these gaps, we will use genomic, genetic, and proteomic methods to profile human macrophages and discover differentially abundant PTMs that are associated with clinical outcomes and/or are Mtb strain dependent. We will then use genetic and cellular strategies to discover the global regulators of anti-microbial macrophage responses to Mtb infection and examine how these genes and their variants regulate macrophage function in the context of Mtb strain variation. Our primary goal is to discover new vulnerabilities between Mtb and macrophages, and thus inform mechanisms of disease heterogeneity, insights into risk stratification for clinical management, and development of effective host directed therapies and vaccines.