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

• 批准号: 9115843
• 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
• 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/9115843
• 关键词: A549; ATP phosphohydrolase; Adjuvant Therapy; Alveolar; Alveolar Macrophages; Antiparasitic Agents; Antitubercular Agents; Antiviral Agents; Autophagocytosis; CD4 Positive T Lymphocytes; CRISPR/Cas technology; Cells; Complex; 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-1; Human; IFITM1 gene; Immune; Individual; Infection; Inflammatory; Integration Host Factors; Interferon Type I; Interferons; Laboratories; Lead; Lung; Mediating; Membrane; Messenger RNA; Molecular; Mycobacterium tuberculosis; Myeloid Cells; Pathway interactions; Patients; Phagosomes; Pharmaceutical Preparations; Phosphoric Monoester Hydrolases; Phosphorylation; Production; Protein Family; RNA helicase A; Regimen; Regulation; Reporting; Risk; Role; STAT3 gene; Signal Transduction; Stress; Syndrome; T-Cell Depletion; TLR2 gene; Testing; Toll-like receptors; Transcriptional Activation; Tretinoin; Tuberculosis; Virulent; Virus Diseases; Virus Replication; Work; antimicrobial drug; base; biological adaptation to stress; co-infection; cytokine; design; eIF-2 Kinase; improved; knock-down; macrophage; member; molecular targeted therapies; monocyte; mortality; mycobacterial; nitazoxanide; novel; overexpression; pathogen; public health relevance; reactivation from latency; reconstitution; research study; response; sensor; small hairpin RNA; small molecule; therapeutic target; treatment duration; tuberculosis treatment; viral RNA

 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) 会增强人类免疫缺陷病毒 (HIV) 的复制和疾病进展，而 HIV 诱导的 CD4+ T 细胞耗竭会显着增加 MTb 的潜伏期重新激活和新 MTb 感染的风险。更好地了解艾滋病毒感染背景下宿主控制潜伏性和活动性结核病的免疫学和细胞机制对于设计结核病/艾滋病毒合并感染的新型治疗方法至关重要。在此，我们提供的初步数据表明，人类单核细胞和单核细胞源性巨噬细胞 (MDM) 的 MTb 感染强烈诱导抗病毒干扰素诱导跨膜 (IFITM) 蛋白家族（IFITM1、2 和 3）的转录。此外，使用 shRNA，我们发现 IFITM1-3 限制人类单核细胞中的 MTb 和 HIV-1 感染。此外，IFITM3 与 MTb 共定位于感染后早期和晚期的内体区室中，并且 IFITM3 的过度表达导致 MTb 感染的单核细胞中内体区室的酸化增强。我们还表明，dead(d)Cas9-KRAB 介导的先天免疫病毒 RNA 传感器、干扰素诱导的双链 RNA 激活蛋白激酶 (PKR) 和视黄酸诱导基因 1 (RIG-I) 的敲低，导致人单核细胞中 MTb 的生长加快。我们对 FDA 批准的小分子药物硝唑尼特 (NTZ) 的研究广泛用于寄生虫性腹泻，该药物先前已被证明可以激活 PKR，诱导应激颗粒的形成，并激活人肺泡上皮细胞中应激反应和抗病毒磷酸酶 GADD34 的转录，这也是由 MTb 诱导的。此外，NTZ 还能抑制 MDM 中的 MTb 和 HIV 感染，并抑制单核细胞中 TLR 介导的静止前病毒 HIV 小基因组的激活。基于这些初步数据，在本提案中，我们将测试以下主要假设：（i）IFITM3是参与骨髓细胞中MTb限制的主要IFITM家族成员，并通过促进v-ATPase招募到分枝杆菌吞噬体导致随后的吞噬体酸化来限制MTb存活和生长； ii) PKR 和 RIG-I 的表达和激活增强了宿主细胞对有毒 MTb 的反应，这种反应是由以下触发的信号启动的： 感染后早期 MTb DNA； iii) NTZ 诱导的应激颗粒形成以及 GADD34 转录本身的激活和 GADD34 介导的 RIG-I 激活都需要 PKR，从而增强对 MTb 和 HIV 的检测和控制； iv) NTZ 通过破坏 STAT3-PKR 基础相互作用并促进 PKR 依赖性自噬来抑制两种病原体的生长。我们期望确定限制 TB/HIV 的新宿主细胞机制，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 是在合并感染的情况下同时调节活动性/潜伏性结核病和艾滋病毒的有吸引力的治疗靶点。我们还预计，我们将确定 NTZ 介导的抗结核/艾滋病毒活性的分子和免疫学相关性，为快速重新利用 NTZ 治疗结核病/艾滋病毒奠定科学基础。