Pseudomonas-Macrophage IL-1β Interactions in Lung Transplant Recipients

肺移植受者中假单胞菌-巨噬细胞 IL-1β 的相互作用

• 批准号: 10606768
• 负责人: Rachel Noel Britton
• 金额: $ 6.95万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10606768
• 关键词: Address; Allografting; Alveolar; Alveolar Macrophages; Bacteria; Bioinformatics; Bronchoalveolar Lavage; CASP1 gene; Categories; Cell Line; Cessation of life; Chronic; Chronic Lung Injury; Clinical; Critical Thinking; Data; Development; Diagnosis; Exposure to; Fellowship; Functional disorder; Future; Human; IL18 gene; IL8 gene; Immune; Immunophenotyping; Impairment; Incidence; Individual; Infection; Inflammasome; Inflammation; Inflammatory; Inflammatory Response; Injury; Innate Immune Response; Interleukin-1 beta; Interleukin-6; Kinetics; Knowledge; Laboratories; Life; Link; Lung; Lung Transplantation; Lung diseases; Macrophage; Measures; Mediating; Mediator; Methods; Modeling; National Research Service Awards; Outcome; Phagocytes; Phenocopy; Phenotype; Play; Population; Preventive; Production; Proteins; Pseudomonas; Pseudomonas aeruginosa; Pulmonary Inflammation; Research; Ribosomal RNA; Risk Factors; Role; Sampling; Scientist; TNF gene; Testing; Transplant Recipients; Transplantation; Upper respiratory tract; bactericide; biobank; career; clinical diagnosis; cytokine; graft failure; host-microbe interactions; immune activation; inhibitor; lung allograft; lung colonization; lung injury; lung microbiome; lung microbiota; microbial; microbiome; microbiome alteration; monocyte; novel; pathogen; pharmacologic; prevent; pulmonary function; research and development; response; sensor; skills; therapeutic target

The objectives of this NRSA individual fellowship are to 1) facilitate the development of research skills necessary for the applicant to become an effective and independent scientist and 2) investigate the mechanisms behind Pseudomonas aeruginosa-induced interleukin-1 beta (IL-1β) production in lung transplantation. Lung transplantation is a life-extending treatment for individuals with end-stage lung diseases; however, long-term outcomes are limited by allograft injury and chronic lung allograft dysfunction (CLAD). While CLAD is defined as a clinical diagnosis with median onset several years after transplantation, the antecedent risk factors may occur months or years prior to diagnosis, supporting a hypothesis that early inflammatory mechanisms could represent important triggers for lung inflammation and injury. Increased bacterial burden, decreased bacterial diversity, and prominence of P. aeruginosa are associated with worsening lung function, graft failure, and CLAD through pathogen-driven inflammatory triggers and impaired innate responses impacting bacterial clearance. The applicant has preliminary data demonstrating that P. aeruginosa upregulates sustained IL-1β production in lung transplant recipient-derived alveolar macrophages compared to other common upper respiratory tract bacteria. Elevated IL-1β has been associated with a decline in lung function and lung inflammation, and chronic lung injury, but few studies have examined the role of pathogen-driven inflammatory triggers on human activation of alveolar macrophage innate immune responses in lung transplantation. This proposal focuses on the mechanisms underlying the interaction between the lung microbiota and its innate sensing by macrophages to address this knowledge gap. We hypothesized that the lung microbiome, particularly those dominated by P. aeruginosa, following transplantation mediates sustained alveolar macrophage proinflammatory responses. To assess the effects of the lung microbiome on macrophages, we used a novel method to isolate lung microbiota from lung transplant recipients for stimulation of macrophages. This proposal uses two approaches to evaluate the impact of P. aeruginosa and Pseudomonas-dominant lung microbiota on immune activation and inflammation: 1) focusing on IL-1β production through activation of the inflammasome using specific canonical sensor proteins and 2) focusing on macrophage immunophenotype. Rigorous immunophenotyping and advanced bioinformatic analysis will be used to elucidate the relationship between P. aeruginosa and inflammasome-activated macrophage IL-1β production in lung transplant recipients. This proposal will add a novel understanding of cellular mechanisms linking microbial lung populations to aberrant innate immune activation, which is critical to developing future therapeutic targets and preventing immune-mediated lung injury. This proposal will provide a rich opportunity for the applicant to establish the laboratory and critical thinking skills necessary for a successful research career.

该NRSA个人奖学金的目标是1）促进必要的研究技能的发展 适当成为有效且独立的科学家和2）研究背后的机制 铜绿假单胞菌在肺移植中产生的白细胞介素-1β（IL-1β）产生。肺 移植是末期肺部疾病患者的延长生命治疗方法。但是，长期 结果受到同种异体损伤和慢性肺同种异体功能障碍（CLAD）的限制。而clad被定义为 移植几年后几年的临床诊断，可能发生前期风险因素 诊断前几个月或几年，支持早期炎症机制可以代表的假设 肺部感染和损伤的重要触发因素。细菌燃烧增加，细菌多样性降低， 铜绿假单胞菌的突出与后悔的肺功能，移植失败和穿越 病原体驱动的炎症触发因素和影响细菌清除率的先天反应受损。这 申请人的初步数据表明，铜绿假单胞菌上调肺中的IL-1β产生 与其他常见的上呼吸道细菌相比，移植受者衍生的肺泡巨噬细胞。 IL-1β升高与肺功能和肺部注射的下降有关，慢性肺 受伤，但很少有研究检查了病原体驱动的炎症触发因素对人类激活的作用 肺巨噬细胞在肺移植中的先天免疫反应。该提议重点是 肺微生物群与其先天感应巨噬细胞之间相互作用的机制 解决此知识差距。我们假设肺微生物组，尤其是由P。 移植后，铜绿可介导持续的肺泡巨噬细胞促炎反应。到 评估肺微生物组对巨噬细胞的影响，我们使用了一种新方法来分离肺微生物群 从肺移植受者刺激巨噬细胞。该建议使用两种方法来评估 铜绿假单胞菌和假单胞菌为主导的肺微生物群对免疫激活和 炎症：1）使用特定规范的炎症小体专注于IL-1β的产生 传感器蛋白和2）专注于巨噬细胞免疫表型。严格的免疫表型和 先进的生物信息学分析将用于阐明铜绿假单胞菌与 肺移植受者中炎症体激活的巨噬细胞IL-1β产生。该建议将增加 对将微生物肺种群与异常先天免疫联系起来的细胞机制的新了解 激活，这对于开发未来的治疗靶标和防止免疫介导的肺损伤至关重要。 该建议将为申请人提供丰富的机会，以建立实验室和批判性思维技能 成功的研究职业所必需的。