Immune control mechanisms of TB latency in the setting of HIV co-infection

HIV合并感染情况下结核潜伏期的免疫控制机制

基本信息

批准号：
9229528
负责人：
ANNE GOLDFELD
金额：
$ 44.25万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9229528
关键词：
A549 Adjuvant Therapy Alveolar Alveolar Macrophages Antiparasitic Agents Antitubercular Agents Antiviral Agents Autophagocytosis CD4 Positive T Lymphocytes CRISPR/Cas technology Cells Complex Cyclic AMP-Dependent Protein Kinases Cyclic GMP Cytoplasmic Granules DNA Data Detection Diarrhea Disease Disease Progression Drug Interactions Drug resistance Drug toxicity Epithelial Cells FDA approved Family member Foundations Gene Expression Genes Genetic Transcription Goals Growth HIV HIV Infections HIV-1 Human IFITM1 gene Immune Immunologics Individual Infection Inflammatory Integral Membrane Protein Integration Host Factors Interferon Type I Interferons Laboratories Lead Lung Mediating Messenger RNA Molecular Mycobacterium tuberculosis Myeloid Cells Pathway interactions Patients Phagosomes Pharmaceutical Preparations Phosphoric Monoester Hydrolases Phosphorylation Production Protein Family Protein Kinase Protein Kinase Interaction RNA helicase A Recruitment Activity Regimen Regulation Reporting Risk Role STAT3 gene Signal Transduction Stat3 protein Stress Syndrome TLR2 gene Testing Toll-like receptors Transcriptional Activation Tretinoin Tuberculosis Virulent Work antimicrobial drug base biological adaptation to stress co-infection cytokine design eIF-2 Kinase experimental study immunoregulation improved knock-down macrophage member molecular targeted therapies monocyte mortality mycobacterial nitazoxanide novel overexpression pathogen protein activation public health relevance reactivation from latency reconstitution response sensor small hairpin RNA small molecule therapeutic target treatment duration tuberculosis treatment vacuolar H+-ATPase viral RNA

项目摘要

DESCRIPTION (provided by applicant) Mycobacterium tuberculosis (MTb) drives enhanced human immunodeficiency virus (HIV) replication and disease progression, while HIV-induced CD4+ T cell depletion dramatically increases MTb reactivation from latency and the risk of de novo MTb infection. Improved understanding of the immunological and cellular mechanisms involved in host control of latent and active TB in the context of HIV infection is critical for the design of novel treatments for TB/HIV co-infection. Here, we present preliminary data demonstrating that transcription of the antiviral interferon-induced transmembrane (IFITM) family of proteins (IFITM1, 2, and 3) is strongly induced by MTb infection of human monocytes and monocyte-derived-macrophages (MDM). Further, using shRNA, we show that IFITM1-3 restrict both MTb and HIV-1 infection in human monocytic cells. In addition, IFITM3 co-localizes with MTb in early and late endosomal compartments post-infection, and overexpression of IFITM3 results in enhanced acidification of endosomal compartments in MTb-infected monocytes. We also show that dead(d)Cas9-KRAB-mediated knock-down of the innate immune viral RNA sensors, interferon-induced, double-stranded RNA-activated protein kinase (PKR), and retinoic acid-inducible gene 1 (RIG-I), leads to greater MTb growth in human monocytic cells. Our studies of the FDA-approved small molecule drug, nitazoxanide (NTZ) widely used in parasitic diarrhea, which has previously been shown to activate PKR, induces the formation of stress granules and activates transcription of the stress response and antiviral phosphatase GADD34 in human alveolar epithelial cells, which is also induced by MTb. Furthermore, NTZ inhibits MTb and HIV infection in MDM, and suppresses TLR-mediated activation of a quiescent proviral HIV minigenome in monocytic cells. Based on these preliminary data, in this proposal we will test the following major hypotheses: (i) IFITM3 is the primary IFITM family member involved in MTb restriction in myeloid cells, and limits MTb survival and growth by promoting recruitment of v-ATPase to the mycobacterial phagosome resulting in subsequent phagosome acidification; ii) PKR and RIG-I expression and activation augment the host cell response to virulent MTb that is initiated by signals triggered by MTb DNA early after infection; iii) PKR is required for NTZ-induced stress granule formation and activation of both GADD34 transcription itself and GADD34-mediated RIG-I activation, which leads to enhanced detection and control of both MTb and HIV; and, iv) NTZ inhibits the growth of both pathogens by disrupting basal STAT3-PKR interactions and promoting PKR-dependent autophagy. We expect to identify novel host cell mechanisms that restrict TB/HIV, including factors that are critical for MTb survival after infection of macrophages, which we anticipate will be attractive therapeutic targets for simultaneous modulation of active/latent TB and HIV in the context of co-infection. We also anticipate that we will identify molecular and immunological correlates of NTZ-mediated anti-TB/HIV activity providing a scientific foundation for rapid repurposing of NTZ for TB/HIV.

描述（由申请人提供）结核分枝杆菌（MTB）驱动人类免疫缺陷病毒（HIV）复制和疾病进展，而HIV诱导的CD4+ T细胞耗竭大大增加了潜伏期的MTB再活化，而DE Novo MTB感染的风险。在HIV感染的背景下，对参与潜在和活性结核病宿主控制的免疫和细胞机制的了解对TB/HIV共同感染的新治疗方法至关重要。在这里，我们介绍了初步数据，表明抗病毒诱导的跨膜（IFITM）蛋白质家族的转录（IFITM1、2和3）是由MTB感染人类单核细胞和单核细胞衍生型巨噬细胞的MTB感染强烈诱导的。此外，使用SHRNA，我们表明IFITM1-3限制了人类单核细胞中的MTB和HIV-1感染。此外，IFITM3在感染前和晚期内体室中与MTB共定位，而IFITM3的过表达导致MTB感染的单核细胞中内体隔室的酸化增强。我们还表明，死去的（D）CAS9-KRAB介导的固有免疫病毒RNA传感器的敲低，干扰素诱导的，双链的RNA激活蛋白激酶（PKR）和视黄酸诱导的基因1（RIG-I）导致人类单细胞细胞中的MTB生长更大。我们对FDA批准的小分子药物（NTZ）广泛用于寄生腹泻中的研究，以前已显示出来激活PKR，诱导应激颗粒的形成，并激活人类Alve Alve Epithelial sirdel alve alve Epithelial，MOTB，MOTB在MORFERALE ELVE ALVE ELVE ELVE ELVE EPITITILIAL，MOTB中，MOTB。 Furthermore, NTZ inhibits MTb and HIV infection in MDM, and suppresses TLR-mediated activation of a quiescent Provided on these preliminary data, in this proposal we will test the following major hypotheses: (i) IFITM3 is the primary IFITM family member involved in MTb restriction in myeloid cells, and limits MTb survival and growth by promoting recruitment of v-ATPase to the分枝杆菌吞噬体导致随后的吞噬体酸化； ii）PKR和RIG-I表达和激活增强了宿主细胞对毒性MTB的反应，该反应是由由由由信号触发的信号引发的感染后MTB DNA； iii）PKR是NTZ诱导的应激颗粒形成和GADD34转录本身和GADD34介导的RIG-I激活所必需的，这导致了MTB和HIV的检测和控制。并且，iv）ntz通过破坏基本的STAT3-PKR相互作用并促进PKR依赖性自噬来抑制两种病原体的生长。我们希望鉴定出限制TB/HIV的新型宿主细胞机制，包括对巨噬细胞感染后MTB存活至关重要的因素，我们预计这将会在共同感染的背景下，可以简单地调节主动/潜在结核和HIV的有吸引力的治疗靶标。我们还预计，我们将确定NTZ介导的抗TB/HIV活性的分子和免疫学相关性，为TB/HIV快速重新利用NTZ的科学基础。