Human macrophage variation & TB pathogenesis

人类巨噬细胞变异

• 批准号: 10459540
• 负责人: Thomas R Hawn
• 金额: $ 49.95万
• 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: 2021-08-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10459540
• 关键词: Address; Adult; Animal Model; Biological Assay; CRISPR/Cas technology; Candidate Disease Gene; Clinical; Clinical Management; Data; Data Set; Development; Disease; Disease Progression; Dissection; Enrollment; Family; Genes; Genetic; Genetic Polymorphism; Genetic Transcription; Genomics; Goals; Heterogeneity; Household; Human; Human Genetics; Immunogenetics; Individual; Interferons; Interleukin-1; Interleukin-1 beta; Knowledge; Meningeal Tuberculosis; Methods; Molecular; Mus; Mycobacterium tuberculosis; Outcome; Pathogenesis; Pathway interactions; Phosphorylation; Post-Translational Protein Processing; Proteins; Proteomics; Pulmonary Tuberculosis; Regulation; Regulator Genes; Resistance; Resistance to infection; Role; Salvelinus; Signal Transduction; Testing; Tuberculosis; Uganda; Vaccines; Variant; Vesicle; Vietnam; antimicrobial; cohort; cytokine; detection method; differential expression; disease heterogeneity; follow-up; gene discovery; genetic association; genetic variant; genome wide association study; in vivo Model; induced pluripotent stem cell; insight; knockout gene; macrophage; monocyte; novel therapeutic intervention; pathogen; protein protein interaction; proteomic signature; resistance mechanism; response; risk stratification; small molecule inhibitor; trafficking; transcriptome sequencing

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.

MTB暴露后临床结局的差异范围从耐药到感染再到传播疾病。 人类的抗药性和传播的遗传和细胞机制在很大程度上是未知的，可能 提供有关新型治疗策略的见解。在越南，我们参加并检查了大量PTB队列 和TBM受试者，对宿主和病原体进行详细的免疫遗传研究，以发现确定剂 疾病进展。在过去20年中，在乌干达的大型家庭联系研究中，我们发现 约9％的亲密家庭接触持续的TST和Interferon-释放测定法（IGRA） 在扩展随访期间为负，并且似乎对传统定义的LTBI（RSTR）具有抵抗力。和 MTB感染的单核细胞的转录和蛋白质组学分析，我们发现了基因和途径 与LTBI相比，在RSTR中富集。在两个数据集中，有些基因都富含 Rab11b，一种参与囊泡运输的基因，在RSTR巨噬细胞中增加了 在人类中。结核病蛋白 - 蛋白质相互作用（PPI）网络。此外，带有细胞GWAS 在MTB感染的巨噬细胞中，我们发现了与IL-1β相关的人类多态性 以前未知基因中调节IL-1的表达。这些发现暗示了新基因和 全局调节人MTB诱导的IL-1β的变体，这是一种促进MTB控制的关键细胞因子 巨噬细胞和鼠体内模型。 RSTRS＆LTBI（乌干达）和 TBM和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 与临床结局密切相关的疾病或巨噬细胞IL-1调节。在一起，这些 数据支持我们的假设，即MTB诱导的巨噬细胞反应受到遗传调节并相关的 有不同的临床结果。但是，我们的知识有很多差距。首先，MTB诱导的后 RST，LTBI和TB疾病个体的巨噬细胞的翻译修饰（PTM）曲线是 未知。其次，MTB诱导的巨噬细胞抗 - 全球人类遗传调节剂（基因和变体） 微生物途径大多未知。第三，人类MTB抗性的分子和细胞机制 传播几乎是完全未知的，包括MTB菌株变异在发病机理中的作用。 为了解决这些差距，我们将使用基因组，遗传和蛋白质组学方法来介绍人类巨噬细胞 并发现与临床结果和/或MTB菌株相关的不同丰富的PTM 依赖。然后，我们将使用遗传和细胞策略来发现抗微生物的全球调节剂 巨噬细胞对MTB感染的反应，并检查这些基因及其变体如何调节巨噬细胞 在MTB应变变化的背景下功能。我们的主要目标是发现MTB之间的新漏洞 和巨噬细胞，从而为疾病异质性的机制提供信息，对风险分层的见解 临床管理，并开发有效的宿主定向疗法和疫苗。