Lung Dysbiosis and Increased Host Susceptibility to Respiratory Pathogens

肺部生态失调和宿主对呼吸道病原体的易感性增加

• 批准号: 10661053
• 负责人: Benjamin G Wu
• 金额: --
• 依托单位: VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10661053
• 关键词: Acetates; Acute; Affect; Airway Disease; Alveolar Macrophages; Antibiotics; Antigens; Bioinformatics; Biological Assay; C57BL/6 Mouse; CD28 gene; CD3 Antigens; Cell Separation; Cell physiology; Cells; Cessation of life; Chronic; Chronic Obstructive Pulmonary Disease; Clinical; Culture-independent methods; Data; Development; Diagnosis; Disease; Ecology; Evaluation; Event; Experimental Designs; Foundations; Frequencies; Future; Gastroesophageal reflux disease; Genus Mycobacterium; Hospitalization; Human; Human body; IL17 gene; Immune; Immune checkpoint inhibitor; Immune response; Immunity; Impairment; Infection; Inflammation; Inflammatory; International; Investigation; Laboratories; Lung; Lung diseases; Malignant neoplasm of lung; Measurement; Measures; Mediating; Mentors; Mentorship; Methodology; Molecular; Mucosal Immunity; Mus; Outcome; Pathogenicity; Pathway Analysis; Pathway interactions; Pattern; Phenotype; Physicians; Pneumonia; Population; Predisposition; Prevotella; Prevotella melaninogenica; Process; Production; Propionates; Proteins; Publications; Pulmonary Inflammation; Recovery; Recurrence; Regulatory T-Lymphocyte; Research; Research Personnel; Resources; Respiratory Mucosa; Respiratory Tract Infections; Risk; Role; Scientist; Specificity; Streptococcus; Streptococcus mitis; Streptococcus pneumoniae; Systems Biology; T cell regulation; T cell response; T-Lymphocyte; TLR4 gene; Techniques; Testing; Training; Translational Research; United States Department of Veterans Affairs; Up-Regulation; Urothelium; Veillonella; Veillonella parvula; Veterans; Volatile Fatty Acids; aspirate; career; checkpoint inhibition; chronic inflammatory lung disease; cohort; dysbiosis; epidemiologic data; exhaustion; experimental study; host microbiota; human subject; immunoregulation; in vivo; infection risk; inflammatory marker; lung microbiota; metabolome; metabolomics; metatranscriptome; metatranscriptomics; microbial; microbial signature; microbiota; microbiota profiles; military veteran; mortality; mouse model; multiple omics; new therapeutic target; next generation sequencing; oral commensal; pathogen; pre-clinical; programmed cell death protein 1; recruit; respiratory challenge; respiratory colonization; respiratory microbiota; respiratory pathogen; response; risk stratification; single-cell RNA sequencing; transcriptomics; trend

Lower airway infections are a significant burden for Veterans. Current therapies commonly involve the use of broad-spectrum antibiotics that lack pathogen specificity and decimates microbial ecology in the human body. The discovery of lung microbiota with the use of next-generation sequencing identified a diverse lower airway microbial community. Our group has shown that the enrichment of the lung microbiota with human oral commensals, such as Streptococcus, Veillonella, and Prevotella, is a hallmark of lower airway dysbiosis associated with increased inflammation and immune exhaustion markers. This dysbiotic signature can be seen in healthy subjects and occurs with higher frequency with chronic inflammatory lung disease such as Chronic Obstructive Pulmonary Disease and lung cancer. In this Veterans Affairs BLR&D CDA2 resubmission, I seek to demonstrate how chronic lung dysbiosis is associated with increased pathogen susceptibility by modulation of the lung immune tone through anaerobic microbial metabolites such as short-chain fatty acids (SCFAs) in a pre- clinical murine model. In our publications, lower airway dysbiosis is associated with increased SCFAs, blunted T cell response to pathogen associated molecular patterns, and increased expression of exhaustion markers and Tregs. My mouse model of induced chronic lower airway dysbiosis and in my preliminary data show increased expression of exhaustion pathways, PD1+ T cells and Tregs in the lung, and increased respiratory pathogen susceptibility in the form of murine mortality and pathogen recovery. The proposed study will use my chronic lung dysbiosis murine model to define the molecular mechanisms leading to the host susceptibility to respiratory pathogens. In Aim 1, I will identify microbiota signatures with the use of metatranscriptome and metabolome approaches present in the chronic murine lower dysbiotic airway, including measurement of SCFA levels and bacterial functions affecting them. I will also test whether SCFAs are sufficient to alter host pathogen susceptibility. In Aim 2, I will identify and test host functional phenotypes of immune exhaustion in my chronic lower airway dysbiosis murine model using fluorescent-activated cell sorting, single-cell RNA-Seq, and assessment of T cell subpopulation function following anti-CD3/CD28 activation. I will also evaluate the role of immune exhaustion on pathogen susceptibility by using checkpoint inhibition during chronic dysbiosis to assess if anti-PD-1 will restore pathogen response. These studies will be performed under the mentorship of Drs. Leopoldo Segal, Xue-Ru Wu, Jun-Chieh Tsay and Jeffrey Weiser. Dr. Segal is my primary mentor and a VA affiliated researcher. He is an expert in systems biology approaches utilizing next-generation sequencing to evaluate airway disease. His expertise in bioinformatic approaches to translational research places him at the forefront of lower airway multi-omic research. My co-mentor is Dr. Xue-Ru Wu who is an expert on urothelial disease and built a career in assessing pathogenic mechanisms that will be highly relevant for my mentorship. Although his expertise is focused on the urothelial tract, his training in experimental design, assessment of molecular mechanisms, and use of murine models will be critical for my scientific training. Dr. Jun-Chieh Tsay is a VA physician scientist with a research focus on lung inflammation in lung cancer. As a co-mentor, he will be a significant resource to assess host inflammation, transcriptomic pathway analysis, and for my career with the VA. In addition, I recruited Dr. Weiser as a co-mentor for his expertise as an international expert in airway mucosal immunity and infection. The mentors and their laboratories, together with the expertise of Veterans Affairs and NYU core laboratories, will provide the optimal setting for the development of my research career. The BLR&D CDA2 will provide the support for my training in the methodologies and techniques necessary to complete the proposed experiments. This proposal will provide the fundamental mechanistic understanding to expand to future investigations involving human cohorts.

下呼吸道感染对退伍军人来说是一个重大负担。目前的治疗方法通常涉及使用 缺乏病原体特异性并破坏人体内微生物生态的广谱抗生素。 通过使用下一代测序发现肺部微生物群，确定了多样化的下呼吸道 微生物群落。我们的小组已经表明，人类口腔可以丰富肺部微生物群 共生菌，如链球菌、韦荣球菌和普雷沃菌，是下呼吸道菌群失调的标志 与炎症和免疫耗竭标志物增加有关。这种失调的特征是可见的 在健康受试者中，慢性炎症性肺病（如慢性炎症）的发生频率更高 阻塞性肺病和肺癌。在退伍军人事务部 BLR&D CDA2 重新提交中，我寻求 通过调节慢性肺生态失调与病原体易感性增加之间的关系 通过厌氧微生物代谢物（例如预-中的短链脂肪酸（SCFA））调节肺部免疫张力 临床小鼠模型。在我们的出版物中，下气道生态失调与 SCFA 增加、减弱有关 T 细胞对病原体相关分子模式的反应，以及耗竭标记表达的增加 和特雷格斯。我的诱导慢性下呼吸道生态失调的小鼠模型和我的初步数据显示 肺中耗竭途径、PD1+ T 细胞和 Tregs 的表达增加，呼吸增加 以小鼠死亡率和病原体恢复形式表现的病原体易感性。 拟议的研究将使用我的慢性肺生态失调小鼠模型来定义分子机制 导致宿主对呼吸道病原体的易感性。在目标 1 中，我将通过以下方式识别微生物群特征： 使用慢性小鼠下呼吸道失调中存在的元转录组和代谢组方法， 包括测量 SCFA 水平和影响它们的细菌功能。我也会测试 SCFA 是否 足以改变宿主病原体的敏感性。在目标 2 中，我将识别并测试宿主功能表型 使用荧光激活细胞分选在我的慢性下气道生态失调小鼠模型中进行免疫衰竭， 单细胞 RNA 测序，以及抗 CD3/CD28 激活后 T 细胞亚群功能的评估。我会 还通过在免疫衰竭过程中使用检查点抑制来评估免疫衰竭对病原体易感性的作用 慢性生态失调，以评估抗 PD-1 药物是否会恢复病原体反应。 这些研究将在博士的指导下进行。莱奥波尔多·西格尔、吴学如、蔡俊杰 和杰弗里·韦瑟。 Segal 博士是我的主要导师，也是 VA 附属研究员。他是系统方面的专家 利用下一代测序来评估气道疾病的生物学方法。他的专业知识 转化研究的生物信息方法使他处于下呼吸道多组学的前沿 研究。我的搭档吴学如医生是尿路上皮疾病专家，并在以下领域建立了职业生涯： 评估与我的指导高度相关的致病机制。虽然他的专业知识是 专注于尿路上皮，他在实验设计、分子机制评估和 小鼠模型的使用对于我的科学训练至关重要。 Jun-Chieh Tsay 博士是退伍军人管理局医师科学家 研究重点是肺癌中的肺部炎症。作为共同导师，他将成为重要资源 评估宿主炎症、转录组通路分析以及我在 VA 的职业生涯。另外，我 聘请Weiser博士作为联合导师，因其作为气道粘膜免疫领域的国际专家的专业知识 和感染。导师及其实验室，以及退伍军人事务部和纽约大学核心的专业知识 实验室，将为我的研究事业的发展提供最佳的环境。 BLR&D CDA2 将 为我完成拟议项目所需的方法和技术方面的培训提供支持 实验。该提案将为扩展至未来提供基本的机制理解 涉及人类队列的调查。