Diversity and function of mononuclear phagocytes in the lung driven by mycobacterium tuberculosis infection

结核分枝杆菌感染驱动的肺部单核吞噬细胞的多样性和功能

• 批准号: 10427702
• 负责人: Beth S Zha
• 金额: $ 17.42万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-02 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10427702
• 关键词: Alveolar Macrophages; Antigens; Automobile Driving; Bacteria; Bioinformatics; Biology; Cell physiology; Cells; Cessation of life; Chronic lung disease; Data; Dendritic Cells; Development; ELF3 gene; Educational workshop; Equipment; Flow Cytometry; Foundations; Genetic Transcription; Genus Mycobacterium; Goals; Growth; Heterogeneity; Homeostasis; Immune; Immune response; Immune system; Immunohistochemistry; Immunology; Infection; Inhalation; Innate Immune Response; International; K-Series Research Career Programs; Laboratories; Leadership; Location; Lung; Lysosomes; Mentors; Mentorship; Mission; Modeling; Mononuclear; Mus; Mycobacterium tuberculosis; Mycobacterium tuberculosis H37Rv; Pathogenicity; Pathway interactions; Phagocytes; Phenotype; Physicians; Play; Population; Regulation; Research; Research Proposals; Resolution; Role; Scientist; Structure of parenchyma of lung; System; T-Lymphocyte; Testing; Time; Training; Tuberculosis; Vaccines; Vacuole; Virulence; Virulence Factors; Work; adaptive immune response; career; cell type; genetic manipulation; improved; infection rate; innovation; lymph nodes; macrophage; monocyte; mouse model; mycobacterial; next generation sequencing; novel; pathogen; programs; recruit; response; single-cell RNA sequencing; symposium; targeted treatment; therapeutic development

Project Summary/Abstract Mycobacterium tuberculosis’s (Mtb) ability to survive and replicate in mononuclear phagocytes (MPs) is central to its pathogenicity and capability to infect a quarter of the world’s population. The laboratory of Dr. Joel Ernst (co-mentor) and others have demonstrated differential Mtb growth and replication between populations of MPs. Yet, the complexity and diversity of MP subsets is only recently being defined at transcriptional resolution and not well described during Mtb infection. It is therefore unclear which subsets are most permissive to Mtb growth and if Mtb directs monocyte differentiation. Using flow cytometry and single cell RNA-sequencing (scRNA-seq) from infected mice, preliminary data demonstrates differential infection rates and responses to Mtb within two recruited macrophage subpopulations. This has directed the central hypothesis that Mtb virulence factors drive differentiation states of recruited MPs in the lung during infection, providing a fertile niche for Mtb growth and replication. The long-term goal of this work is to delineate the relationship between Mtb and MPs in the context of a recent revolution in understanding lung immune cell heterogeneity, enabling more effective host-directed therapies. Utilizing fluorescently expressing Mtb, flow cytometry, and single cell-RNA sequencing in mice infected with a laboratory strain of Mtb (H37Rv), and a strain lacking a functional ESX-1 secretion system, specific Aim 1 will test the hypothesis that an Mtb virulence system drives differentiation of monocytes to specific macrophage subsets. With use of flow cytometry, immunohistochemistry, and ex vivo infection models, specific Aim 2 will test the hypothesis that recruited macrophage subsets have differential ability to restrict Mtb replication. The training plan is focused to develop expertise in immunologic responses to mycobacterium, mononuclear phagocyte and lung milieu biology, genetic manipulation of mycobacterium, scRNA-seq and bioinformatics, leadership, and responsible research practices. Training will include didactic courses, workshops, seminars, local and international conferences. The primary mentor and co-mentors are Dr. Oren Rosenberg and Dr. Joel Ernst, both successful physician-scientists with combined expertise in immunology, Mtb biology, and host- pathogen interactions. An overarching team of mentors also include a scientific advisor in bioinformatics, collaborator in next generation sequencing, and accomplished physician-scientist career mentors. Dr. Zha has laboratory and office space at UCSF Mission Bay, with shared space in a BSL2 and BSL3 laboratories, with access to equipment and mouse models necessary for this research. This Career Development Award application has an innovative research proposal, rigorous training plan, mentorship of leaders in mycobacterial immunology and biology, team of physician-scientist advisors with a track record of transitioning scientists to independence, and an exceptional candidate driven toward an independent career in mycobacterial-host interaction and chronic lung disease.

项目概要/摘要 结核分枝杆菌 (Mtb) 在单核吞噬细胞 (MP) 中的生存和复制能力是 其致病性和感染世界四分之一人口的能力至关重要。乔尔博士的实验室 恩斯特（共同导师）和其他人已经证明了 Mtb 的生长和复制在不同人群之间存在差异。 议员们。然而，MP 子集的复杂性和多样性最近才在转录分辨率上被定义 在 Mtb 感染期间没有得到很好的描述。因此，尚不清楚哪些子集对 Mtb 最宽容 生长以及 Mtb 是否指导单核细胞分化。使用流式细胞术和单细胞 RNA 测序 （scRNA-seq）来自受感染的小鼠，初步数据显示了不同的感染率和对 Mtb 的反应 在两个招募的巨噬细胞亚群内。这指导了中心假设：结核分枝杆菌毒力 感染期间肺部招募的MPs的分化状态由多种因素驱动，为结核分枝杆菌提供了肥沃的生态位 生长和复制。这项工作的长期目标是描绘 Mtb 和 MPs 之间的关系 在最近理解肺免疫细胞异质性的革命背景下，能够更有效地 宿主导向疗法。利用荧光表达 Mtb、流式细胞术和单细胞 RNA 测序 在感染 Mtb 实验室菌株 (H37Rv) 和缺乏功能性 ESX-1 分泌系统的菌株的小鼠中， 具体目标 1 将检验以下假设：Mtb 毒力系统驱动单核细胞分化为特定的 巨噬细胞亚群。通过使用流式细胞术、免疫组织化学和离体感染模型，特异性 目标 2 将检验以下假设：招募的巨噬细胞亚群具有不同的限制 Mtb 的能力 复制。 该培训计划的重点是发展对分枝杆菌、单核细胞的免疫反应方面的专业知识。 吞噬细胞和肺环境生物学、分枝杆菌基因操作、scRNA-seq 和生物信息学， 领导力和负责任的研究实践。培训将包括教学课程、讲习班、研讨会、 本地和国际会议。主要导师和共同导师是 Oren Rosenberg 博士和 Joel 博士 恩斯特（Ernst）都是成功的医师科学家，在免疫学、结核分枝杆菌生物学和宿主方面拥有综合专业知识。 病原体相互作用。一个总体导师团队还包括一名生物信息学科学顾问， 下一代测序领域的合作者，以及卓有成就的医师科学家职业导师。查博士有 UCSF Mission Bay 的实验室和办公空间，在 BSL2 和 BSL3 实验室中共享空间， 获得本研究所需的设备和小鼠模型。本次职业发展奖 应用程序有创新的研究计划、严格的培训计划、分枝杆菌领导者的指导 免疫学和生物学，由医师科学家顾问组成的团队，拥有将科学家转变为 独立性，以及一位在分枝杆菌宿主中走向独立职业生涯的杰出候选人 相互作用和慢性肺部疾病。