Macrophage and monocyte TBK1 function in host defense against influenza

项目摘要 本K 08指导临床科学家职业奖申请的目的是促进以下方面的发展： 基本技能，这将使候选人成为一个学术物理学家，科学家。候选人及其 导师们设计了一个培训计划，其中将包括一个严格的研究组成部分，沿着教学 指导建立成功的职业发展所需的思维过程和原则。 通过课程和实践经验，候选人将对 流感发病机制和宿主对流感病毒的免疫反应的复杂免疫机制 感染通过他在重症监护室的持续临床工作，候选人将获得 有机会告知他的基础研究计划，以及指导该工作的临床研究的翻译 和实践流感和其他呼吸道病毒导致数千例急性呼吸窘迫 急性呼吸窘迫综合征（ARDS）仅在美国每年发生一次，是发病率和死亡率的重要驱动因素 国际吧尽管有抗流感病毒药物，但仍然迫切需要宿主- 限制器官损伤的定向治疗方法，可用于多种病毒性疾病。初始 对病毒感染的应答集中在先天免疫系统，包括1型干扰素（IFN）表达 从受感染的细胞和响应细胞如巨噬细胞和单核细胞。多种病毒和炎症 输入集中于信号激酶，如TANK结合激酶1（TBK 1），这些蛋白质调节 以高度细胞类型特异性的方式抑制免疫和炎症。TBK 1的免疫促进作用 除1型外，IFN的表达仍高度不确定。候选人的数据表明，TBK 1 巨噬细胞和单核细胞的功能促进炎性巨噬细胞在肺中的积聚， 删除髓系TBK 1可以降低宿主发病率，同时保持病毒控制。TBK 1的新发现 在巨噬细胞中的功能表明TBK 1轴是炎症性肺病中可能的药物靶点。 该提案定义了TBK 1对肺中单核细胞和巨噬细胞驱动的炎症的贡献 感染的目标是确定可操作的机制，可以操纵主机定向 治疗学具体目标是利用体内和体外技术研究单核细胞和巨噬细胞 功能和先天免疫信号传导的分子方面。在目标1中，我们将确定哪些单核细胞 亚型和巨噬细胞亚群需要TBK 1进入流感感染的肺部，并检查 TBK 1功能的转录组学后果。在目标2中，我们将使用小鼠模型来确定上游 在流感感染中激活巨噬细胞TBK 1的信号。最后，在目标3中，我们将使用生物化学和 蛋白质组学工具来检查病毒和趋化因子驱动的TBK 1激活的信号转导结果， 单核细胞和巨噬细胞。这些研究将探索有关TBK 1功能的新观察结果， 最终目标是改善这种常见致命疾病的患者结局。

Project Summary The objective of this K08 Mentored Clinical Scientist Career Award Application is to facilitate development of essential skills that will allow the candidate to become an academic physician-scientist. The candidate and his mentor have designed a training plan that will include a rigorous research component along with didactic instruction to establish the thought processes and principles necessary for successful career development. Through coursework and practical experience, the candidate will develop a deeper understanding of the complex immune mechanisms that underlie influenza pathogenesis and the host immune response to infection. Through his continued clinical work in the medical intensive care unit, the candidate will have the opportunity to inform his basic research program as well as guide the translation of that work to clinical studies and practice. Influenza and other respiratory viruses cause thousands of cases of Acute Respiratory Distress Syndrome (ARDS) yearly in the United States alone and are a substantial driver of morbidity and mortality worldwide. Despite the availability of antiviral medications for influenza, there remains an urgent need for host- directed therapeutics that limit organ damage and can be deployed in a variety of viral illnesses. The initial response to viral infection centers on the innate immune system including type 1 interferon (IFN) expression from infected cells and responding cells such as macrophages and monocytes. Multiple viral and inflammatory inputs converge on signaling kinases such as TANK-binding kinase 1 (TBK1), and these proteins regulate immunity and inflammation in a highly cell type-specific manner. The function of TBK1 in promoting immunity apart from type 1 IFN expression remains highly undefined. The candidate's data demonstrate that TBK1 function in macrophages and monocytes promote accumulation of inflammatory macrophages in the lung, and that deleting myeloid TBK1 lessens host morbidity while preserving viral control. This novel finding of TBK1 function in macrophages points to the TBK1 axis as a possible druggable target in inflammatory lung disease. This proposal defines the contribution of TBK1 to monocyte- and macrophage-driven inflammation in lung infections with the goal of identifying actionable mechanisms that can be manipulated for host-directed therapeutics. The specific aims will utilize in vivo and in vitro techniques to study monocyte and macrophage function and the molecular aspects of innate immune signaling. In Aim 1, we will determine which monocyte subtype and macrophage subpopulation require TBK1 to enter the lung in influenza infection and examine the transcriptomic consequences of TBK1 function. In Aim 2, we will use mouse models to determine the upstream signals that activate macrophage TBK1 in influenza infection. Finally, in Aim 3 we will use biochemical and proteomic tools to examine the signaling consequences of virus- and chemokine-driven TBK1 activation in monocytes and macrophages. These studies will explore new observations concerning TBK1 function with the ultimate goal of improving patient outcomes in this frequently fatal disease.