Elucidating the path to type I IFNs in TB infection

阐明结核感染中 I 型干扰素的途径

• 批准号: 10592443
• 负责人: Amy K Barczak
• 金额: $ 59.39万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10592443
• 关键词: Antitubercular Agents; Bacteria; CRISPR/Cas technology; Cause of Death; Cells; Clinical Treatment; Complex; Data; Development; Dimensions; Disease; Eicosanoid Production; Enzyme-Linked Immunosorbent Assay; Epidemic; Equilibrium; Event; Future; Gene Expression Profile; Gene Expression Profiling; Genetic; Genetic Transcription; Growth; Histopathology; Human; Immune; Immune response; Immunologic Receptors; Infection; Infection Control; Inflammation; Inflammatory Response; Inhalation; Innate Immune Response; Interferon Type I; Interferons; Interleukin-1; Knowledge; Link; Lipids; Literature; Lung; Macrophage; Measures; Microscopy; Mitochondria; Modeling; Molecular; Mycobacterium tuberculosis; Outcome; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Play; Production; Productivity; Role; Sampling; Shapes; Signal Transduction; Testing; Therapeutic; Tissues; Tuberculosis; Western Blotting; Whole Organism; Work; arm; biological adaptation to stress; cytokine; endoplasmic reticulum stress; human disease; improved; in vivo; individual response; innate immune pathways; interest; metabolomics; mouse model; novel; pathogen; pathogenic bacteria; recruit; response; small molecule; small molecule inhibitor; therapeutic target; tool; tuberculosis treatment; uptake

Tuberculosis (TB) is the leading cause of death from infection globally. Our knowledge of the cellular and molecular events that link inhalation of the causative bacterium, Mycobacterium tuberculosis (Mtb), with either clearance or productive infection remains limited. The type I interferon (IFN) response is among the first innate immune responses triggered in Mtb-infected macrophages. Growing evidence suggests that type I IFNs, and specifically cross-talk between the type I IFN response and the IL-1 axis, drive pathogenesis in TB. However, our understanding of the cascade of events that initiate the type I IFN response in Mtb-infected host cells is incomplete. The literature increasingly supports a model in which mitochondrial damage is a key driver of type I IFN production in Mtb-infected macrophages. In preliminary work, we have uncovered a set of previously unappreciated additional cellular pathways that contribute to Mtb-induced type I IFNs, including the ER stress response (ESR), lipid droplet (LD) formation, and eicosanoid production. In the proposed work, we will build upon our preliminary results to define molecular relationships between ER stress, LD formation, eicosanoid production, and mitochondrial damage in type I IFN response to Mtb. We will then use a murine model of infection to test the impact of modulating the ESR on infection outcomes. In Aim 1, we will use CRISPR technology to build genetic tools to study the pathways of interest. Using these tools and small molecule inhibitors, we will then test links between arms of the ESR, LD formation, eicosanoid production, and type I IFNs. To more completely characterize the role of the ESR and individual response pathways in the macrophage response to Mtb, we will additionally perform multiplexed cytokine analysis, transcriptional profiling, and metabolomics using our genetic and small molecule tools that perturb the ESR. In Aim 2, we will test which of the identified contributors to type I IFNs drive mitochondrial damage. In Aim 3, we will use small molecule inhibitors in two murine models of TB infection to determine how modulating the ESR in the context of TB infection changes bacterial burden, immune cell recruitment to the lung compartment, histopathology, cytokine responses, and the transcriptional response. Upon achieving our aims, we anticipate having developed a new, more complex model for induction of type I IFNs in Mtb-infected macrophages. Further, we anticipate having determined how the ESR shapes the macrophage response to Mtb infection and contributes to infection outcomes in vivo. We anticipate these results will ultimately inform the development of novel host directed therapies for TB.

结核病（TB）是全球感染死亡的主要原因。我们对细胞和 将吸入致病菌结核分枝杆菌（Mtb）与以下任一种疾病联系起来的分子事件： 清除或生产性感染仍然有限。I型干扰素（IFN）反应是第一个先天性的 在结核杆菌感染的巨噬细胞中引发的免疫反应。越来越多的证据表明，I型干扰素， 特别是I型IFN应答和IL-1轴之间的相互作用，驱动TB的发病机制。然而，在这方面， 我们对在结核分枝杆菌感染的宿主细胞中启动I型IFN应答的级联反应的理解是， 不完整文献越来越多地支持一种模型，其中线粒体损伤是I型糖尿病的关键驱动因素。 Mtb感染的巨噬细胞中的IFN产生。在初步工作中，我们发现了一套以前 未被重视的额外的细胞途径，有助于结核分枝杆菌诱导的I型干扰素，包括ER应激 反应（ESR）、脂滴（LD）形成和类花生酸产生。在拟议的工作中，我们将建立 根据我们的初步结果来定义ER应激、LD形成、类二十烷酸之间的分子关系 生产，和线粒体损伤的I型干扰素响应结核分枝杆菌。然后，我们将使用小鼠模型， 感染，以测试调节ESR对感染结果的影响。在目标1中，我们将使用CRISPR 技术来构建遗传工具来研究感兴趣的途径。使用这些工具和小分子 抑制剂，然后我们将测试ESR，LD形成，类花生酸生产和I型之间的联系 干扰素。为了更全面地描述ESR和个体反应途径在 巨噬细胞对结核分枝杆菌的反应，我们将另外进行多重细胞因子分析，转录 分析和代谢组学使用我们的遗传和小分子工具，扰乱ESR。在目标2中，我们将 测试哪些已确定的I型干扰素的贡献者驱动线粒体损伤。在目标3中，我们将使用小 分子抑制剂在两个小鼠模型的结核病感染，以确定如何调节ESR的背景下， 结核病感染改变了细菌负荷，免疫细胞募集到肺室，组织病理学， 细胞因子反应和转录反应。在实现我们的目标后，我们预计 开发了一种新的、更复杂的模型，用于在结核分枝杆菌感染的巨噬细胞中诱导I型IFN。我们还 预期已经确定ESR如何塑造巨噬细胞对Mtb感染的反应，并有助于 与体内感染结果的关系。我们预计这些结果将最终为新型宿主的开发提供信息 结核病的定向治疗。