Unfolded Protein Response Activation as a Determinant of the Host Response to M. tuberculosis

据估计，每年约有1000万人患上结核病。即使成功治愈了 抗生素，结核病患者经常因感染相关而导致永久性肺功能受损。 发炎。宿主指导的限制非生产性炎症的治疗是一种有希望的改善工具 疾病后果。这些疗法的发展因观察到的异质性程度而受到阻碍。 对结核分枝杆菌感染的个体反应。为了克服这一限制，必须利用新的方法来 理解这些不同结果背后的途径。申请人建议使用基因上的 近交系重组小鼠品系的不同协作杂交(CC)小组特征 潜在的变量对感染的反应。利用CC创始菌株多样性的初步研究 揭示了未折叠蛋白反应(UPR)活性的变异性可能是免疫异质性的基础 对结核分枝杆菌感染的反应。UPR是一种确保内质网内稳态和 由三个不同的信号通路组成，它们启动互补反应。这些人的活动 通路受多种细胞应激、内环境平衡和病原体感知通路的影响，强调 UPR被认为是许多潜在寄主变异来源下游的汇合点。这项建议 目的：详细说明UPR活性在结核分枝杆菌髓系反应中的作用，并确定 针对个别UPR通路进行治疗干预。目标1中提出的实验将 确定感染期间UPR活性的下游相关性，并评估UPR途径的贡献 利用单细胞RNAseq技术实现转录反应的异质性。目标2中描述的研究 将描述UPR途径的化学和遗传抑制对巨噬细胞功能的影响 在结核分枝杆菌感染期间。AIM 3将评估髓系UPR途径活性对疾病进展的影响 在肺部结核杆菌感染后，使用基于遗传学和基于抑制剂的方法相结合的方法。一起， 这些研究将提供关于免疫反应异质性和普遍定期审议的贡献的见解。 结核分枝杆菌感染结局的途径。申请者将作为Sassetti的一部分进行拟议的研究 马萨诸塞大学陈医学院的研究小组。这个环境将为他们提供 拥有完成研究所需的设施，并由一个合作的科学家小组提供投入 相关领域的专业知识，包括免疫学和细菌学。完成建议的研究后， 应聘者将获得各种实验技术的经验，包括单细胞RNAseq和Flow 细胞学。此外，通过建议书中详细说明的培训，申请者将发展专业人员 以及作为一名独立研究员取得成功所必需的沟通技能。总之，这些研究和 本申请中建议的培训将为申请者提供运营 独立研究计划。

It is estimated that roughly 10 million people fall ill with TB annually. Even when successfully cured with antibiotics, TB patients often suffer from permanently impaired lung function resulting from infection associated inflammation. Host directed therapies that limit nonproductive inflammation represent a promising tool to improve disease outcomes. Development of these therapies has been hindered by the extent of heterogeneity observed in individual responses to Mtb infection. To overcome this limitation, novel approaches must be utilized to understand the pathways underlying these variable outcomes. The applicant proposes the use of the genetically diverse collaborative cross (CC) panel of inbred recombinant mouse strains to characterize the features underlying variable responses to infection. Preliminary studies leveraging the diversity of the CC founder strains revealed that variability in unfolded protein response (UPR) activity may underlie the heterogeneity in immune responses to Mtb infection. The UPR is a mechanism for ensuring endoplasmic reticulum homeostasis and consists of three distinct signaling pathways that initiate complementary responses. The activity of these pathways is influenced by a variety of cell stress, homeostatic, and pathogen sensing pathways, emphasizing the UPRs identity as a convergence point downstream of many potential sources of host variation. This proposal aims to detail the contribution of UPR activity to myeloid responses to Mtb and to determine the feasibility of targeting the individual UPR pathways for therapeutic intervention. The experiments proposed in aim 1 will identify downstream correlates of UPR activity during infection and assess the contribution of the UPR pathways to transcriptional response heterogeneity using single cell RNAseq technology. The studies described in aim 2 will characterize the consequences of chemical and genetic inhibition of UPR pathways on macrophage functions during Mtb infection. Aim 3 will assess the impact of myeloid UPR pathway activity on disease progression following pulmonary Mtb infection using a combination of genetics- and inhibitor- based approaches. Together, these studies will provide insights regarding immune response heterogeneity and the contribution of the UPR pathways to Mtb infection outcomes. The applicant will carry out the proposed studies as part of the Sassetti research group at the University of Massachusetts Chan Medical School. This environment will provide them with the facilities necessary to complete their studies, and input from a collaborative group of scientists with expertise in relevant fields including immunology and bacteriology. By completing the proposed studies, the applicant will gain experience in a variety of experimental techniques, including single cell RNAseq, and flow cytometry. Additionally, through the training detailed in the proposal, the applicant will develop the professional and communication skills necessary for success as an independent researcher. Together, the studies and training proposed within this application will provide the applicant with the skills and experience to run an independent research program.