Elucidating the path to type I IFNs in TB infection

结核病是全球主要的感染致死原因。我们对细胞和细胞的知识 与吸入引起结核分枝杆菌(Mtb)有关的分子事件 清除或生产性感染仍然有限。I型干扰素(干扰素)反应是第一种先天反应。 在结核杆菌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，以及 尤其是I型干扰素反应和IL-1轴之间的串扰，推动了结核病的发病。然而， 我们对在结核分枝杆菌感染的宿主细胞中启动I型干扰素反应的一连串事件的理解是 不完整。文献越来越多地支持线粒体损伤是I型的关键驱动因素的模型 结核分枝杆菌感染的巨噬细胞产生干扰素。在前期工作中，我们发现了一组以前的 未被认可的促进结核分枝杆菌诱导的I型干扰素的细胞通路，包括内质网应激 反应(ESR)、脂滴(LD)的形成和二十烷类化合物的产生。在拟议的工作中，我们将建立 根据我们的初步结果，确定了内质网应激、LD形成、二十烷类化合物之间的分子关系 Mtb对I型干扰素的反应中的产生和线粒体损伤。然后我们将使用一种小鼠模型 以测试调节血沉对感染结局的影响。在目标1中，我们将使用CRISPR 构建基因工具来研究感兴趣的途径的技术。使用这些工具和小分子 抑制剂，然后我们将测试ESR的臂之间的联系，LD的形成，二十烷类化合物的产生，和I型 IFN。为了更全面地描述血沉和个体反应途径在 巨噬细胞对结核分枝杆菌的反应，我们还将进行多重细胞因子分析，转录 使用我们的基因和小分子工具扰乱ESR的简档分析和代谢组学。在目标2中，我们将 测试哪些已识别的I型IFN的贡献者推动线粒体损伤。在目标3中，我们将使用Small 两种结核病感染小鼠模型中的分子抑制物以确定如何在环境中调节ESR 结核病感染会改变细菌负荷、免疫细胞向肺间室的募集、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，我们预计会有 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。此外，我们 预期已确定血沉如何塑造巨噬细胞对结核分枝杆菌感染的反应并做出贡献 体内的感染结果。我们期待这些结果最终将为新型主持人的发展提供参考 结核病的定向治疗。

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.