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.

项目摘要/摘要 结核分枝杆菌（MTB）在单核吞噬细胞（MPS）中生存和复制的能力为 其致病性和能力感染世界四分之一人口的核心。乔尔博士的实验室 恩斯特（Co-intor）和其他人表现出了不同的MTB增长和复制 国会议员。然而，直到最近才在转录分辨率下定义MP子集的复杂性和多样性 并且在MTB感染期间不太描述。因此，尚不清楚哪些子集对MTB最允许 生长，如果MTB指导单核细胞分化。使用流式细胞术和单细胞RNA序列 （SCRNA-SEQ）来自感染小鼠，初步数据表明感染率差异和对MTB的反应 在两个招募的巨噬细胞亚群中。这指示了MTB病毒的中心假设 因素驱动感染期间肺部招募的MP的分化状态，为MTB提供肥沃的利基市场 增长和复制。这项工作的长期目标是描述MTB与MPS之间的关系 在最近了解肺免疫细胞异质性的革命的背景下，使得更有效 宿主定向疗法。利用荧光表达MTB，流式细胞术和单细胞-RNA测序 在感染了MTB（H37RV）和缺乏功能性ESX-1分泌系统的菌株的小鼠中， 特定目标1将检验以下假设：MTB病毒系统将单核细胞分化为特定 巨噬细胞子集。使用流式细胞术，免疫组织化学和离体感染模型，特定 AIM 2将检验以下假设，即招募的巨噬细胞子集具有限制MTB的差异能力 复制。 该培训计划的重点是发展对分枝杆菌的免疫学反应的专业知识 吞噬细胞和肺部环境生物学，分枝杆菌的遗传操纵，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，半小伙子， 地方和国际会议。主要的导师和联合给人员是Oren Rosenberg博士和Joel博士 恩斯特（Ernst 病原体相互作用。一个总体导师团队还包括生物信息学科学顾问， 下一代测序的合作者，并完成了身体科学的职业导师。 Zha博士有 UCSF Mission Bay的实验室和办公空间，在BSL2和BSL3实验室中具有共享空间，并带有 访问此研究所需的设备和鼠标模型。这个职业发展奖 应用程序具有创新的研究建议，严格的培训计划，分生不清的领导者的指导 免疫学和生物学，身体科学家顾问团队，其往绩将科学家过渡到 独立性，以及在分枝杆菌主持人中朝着独立职业的出色候选人开车 相互作用和慢性肺部疾病。