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

基本信息

批准号：
10378549
负责人：
Amy K Barczak
金额：
$ 60.73万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
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/10378549
关键词：
Antitubercular Agents Bacteria CRISPR/Cas technology Cause of Death Cells Clinical Treatment Complex Data Development 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 Measures Microscopy Mitochondria Modeling Molecular Mycobacterium tuberculosis Outcome Pathogenesis Pathogenicity Pathway interactions Patients Play Production 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 macrophage metabolomics mouse model novel pathogen pathogenic bacteria recruit response small molecule small molecule inhibitor therapeutic target tool tuberculosis treatment uptake

项目摘要

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。我们对细胞和细胞的了解连接吸入致病细菌，结核分枝杆菌（MTB）的分子事件，清除或生产感染仍然有限。 I型干扰素（IFN）响应是第一个先天的响应在MTB感染的巨噬细胞中触发的免疫反应。越来越多的证据表明，I型IFN和特定于I型IFN响应和IL-1轴之间的横断性，在TB中驱动发病机理。然而，我们对启动MTB感染宿主细胞中I型IFN响应的级联事件的级联的理解是不完整。文献越来越支持一个模型，在该模型中，线粒体损伤是I型的关键驱动力在MTB感染的巨噬细胞中产生IFN。在初步工作中，我们已经发现了一组以前的未欣赏导致MTB诱导的I型IFN的其他其他细胞途径，包括ER应力反应（ESR），脂肪液滴（LD）形成和类花生酸的产生。在拟议的工作中，我们将建造根据我们的初步结果，定义了ER应力，LD形成，类eicosanoid之间的分子关系 I型IFN对MTB的响应中的生产和线粒体损伤。然后，我们将使用一个鼠模型感染以测试调节ESR对感染结果的影响。在AIM 1中，我们将使用CRISPR 建立遗传工具来研究感兴趣的途径的技术。使用这些工具和小分子然后，我们将测试ESR，LD形成，类类生产和I型的臂之间的链接 IFNS。更彻底地表征了ESR和个体反应途径在巨噬细胞对MTB的反应，我们还将执行多路复用细胞因子分析，转录使用我们的遗传和小分子工具进行分析和代谢组学，这些工具会扰动ESR。在AIM 2中，我们将测试哪个确定的I型IFN的贡献者驱动线粒体损伤。在AIM 3中，我们将使用小 TB感染的两个鼠模型中的分子抑制剂，以确定如何在上下文中调节ESR 结核病感染会改变细菌负担，免疫细胞募集到肺部室，组织病理学，细胞因子反应和转录反应。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MTB感染的巨噬细胞中诱导I型IFN。此外，我们预计已经确定了ESR如何塑造巨噬细胞对MTB感染的反应并贡献在体内感染结果。我们预计这些结果最终将告知新型主机的发展 TB的定向疗法。