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

据估计，每年约有1000万人患结核病。即使成功治愈， 由于抗生素的使用，结核病患者往往会因感染相关疾病而导致肺功能永久受损。 炎症限制非生产性炎症的宿主导向疗法代表了改善炎症的有希望的工具。 疾病结果。这些疗法的发展一直受到所观察到的异质性程度的阻碍 对结核病感染的个体反应。为了克服这一限制，必须利用新的方法来 了解这些变量结果背后的途径。申请人建议使用基因 近交系重组小鼠品系的多样性协作杂交（CC）组以表征特征 对感染的潜在可变反应。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Mtb感染的反应。UPR是确保内质网稳态的机制， 由三个不同的信号通路组成，它们启动互补反应。这些活动 途径受到多种细胞应激、稳态和病原体感应途径的影响， UPR是许多潜在宿主变异源下游的汇聚点。这项建议 目的是详细说明UPR活性对Mtb骨髓反应的贡献，并确定 针对单个UPR途径进行治疗干预。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 使用单细胞RNAseq技术的转录应答异质性。目标2中描述的研究 将描述化学和遗传抑制UPR途径对巨噬细胞功能的影响 在结核病感染期间。目的3将评估髓系UPR通路活性对疾病进展的影响 肺结核感染后使用遗传学和抑制剂为基础的方法相结合。在一起， 这些研究将提供有关免疫反应异质性和普遍定期审议贡献的见解 Mtb感染的结果。申请人将进行拟议的研究，作为Sassetti的一部分。 马萨诸塞州陈医学院的研究小组。这种环境将为他们提供 与必要的设施，以完成他们的研究，并从一个合作小组的科学家的投入， 在相关领域的专业知识，包括免疫学和细菌学。通过完成建议的研究， 申请人将获得各种实验技术的经验，包括单细胞RNAseq和流式细胞术。 细胞仪此外，通过计划书中详细介绍的培训，申请人将发展专业技能， 以及作为独立研究人员取得成功所必需的沟通技巧。总之，研究和 本申请中建议的培训将为申请人提供运行 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.