Selective modulation of neutrophils in critical illness

危重疾病中中性粒细胞的选择性调节

• 批准号: 9750019
• 负责人: CRAIG THOMAS LEFORT
• 金额: $ 40.25万
• 依托单位: RHODE ISLAND HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-05 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9750019
• 关键词: Address; Bacterial Infections; Biological; Biology; Blood Circulation; CRISPR/Cas technology; Coupled; Critical Illness; Disease; Genetic; Genetic Screening; Goals; Health; Hemorrhagic Shock; Hospitals; Host Defense; Human; Immune; Immune System Diseases; Immune system; Immunity; Infection; Injury; Integrins; Knowledge; Leukocytes; Life; Liquid substance; Lung; Mediating; Modeling; Modification; Molecular; Nature; Neutrophil Infiltration; Operative Surgical Procedures; Organism; Phagocytosis; Phenotype; Plasma; Predisposition; Research; Respiratory Tract Infections; Role; Secondary to; Shock; Tissues; Trauma; Trauma patient; Traumatic injury; Work; antimicrobial; cytotoxic; fighting; innovation; mortality; mouse model; neutrophil; pathogenic bacteria; pathogenic fungus; programs; recruit; response; secondary infection

Abstract Critical illness describes the life-threatening state caused by immune dysfunction that can occur following traumatic injury, shock, burn or major surgery. Trauma-induced immune incompetence enhances susceptibility to secondary infections that are a significant cause of mortality in hospital ICUs. Neutrophils, the most abundant leukocyte in the circulation, are critical for host defense against bacterial and fungal pathogens, but they also induce bystander tissue injury due to the non-specific and cytotoxic nature of their antimicrobial arsenal. The effects of trauma on neutrophils contribute to immune dysfunction and include a significant deficit in neutrophil capacity to find and fight infection, coupled with a primed phenotype associated with inducing tissue damage. The long-term goals of my research program are to understand mechanisms of neutrophil recruitment and effector function in health and disease. This MIRA application describes our synergistic approaches to identifying and understanding mechanisms of neutrophil pathobiology during critical illness. First, we have established an innovative approach to derive neutrophils ex vivo that circumvents the technical barriers to their genetic modification. This enables us to perform multiplexed forward genetic screening using CRISPR-Cas9 in neutrophils, and thereby identify new mechanistic aspects of their recruitment (e.g., integrin activation) and antimicrobial function (e.g., phagocytosis). Second, we have established a mouse model of critical illness by respiratory infection secondary to hemorrhagic shock. As the lungs are particularly susceptible to infection secondary to trauma and are also prone to neutrophil-mediated injury, this model provides a means to evaluate new targets/strategies for promoting selective aspects of neutrophil function to enhance host defense without exacerbating tissue damage. Finally, we will apply human trauma patient material (blood, plasma, lung fluid) to these approaches to probe disease-specific mechanisms of the neutrophil response. Since our studies will interrogate fundamental biological mechanisms, such as integrin activation, this work will also have broad impact on multiple fields. Together, these integrated approaches will address major knowledge gaps in neutrophil biology by spanning the molecular, cellular and organism/disease levels of experimentation.

抽象的 危重疾病描述了可能发生的由免疫功能障碍引起的危及生命的状态 外伤、休克、烧伤或重大手术后。创伤引起的免疫功能低下会增强 继发感染的易感性是医院 ICU 死亡的一个重要原因。中性粒细胞， 循环中最丰富的白细胞，对于宿主防御细菌和真菌病原体至关重要， 但由于其抗菌剂的非特异性和细胞毒性，它们也会引起旁观者组织损伤 兵工厂。创伤对中性粒细胞的影响会导致免疫功能障碍，并包括严重的缺陷 中性粒细胞发现和抵抗感染的能力，加上与诱导相关的启动表型 组织损伤。我的研究计划的长期目标是了解中性粒细胞的机制 健康和疾病中的募集和效应器功能。此 MIRA 应用程序描述了我们的协同作用 识别和理解危重病期间中性粒细胞病理学机制的方法。 首先，我们建立了一种创新的方法来体外获取中性粒细胞，从而规避了技术 其基因改造的障碍。这使我们能够使用以下方法进行多重正向遗传筛选 中性粒细胞中的 CRISPR-Cas9，从而识别其募集的新机制（例如，整合素 激活）和抗菌功能（例如吞噬作用）。其次，我们建立了小鼠模型 因呼吸道感染继发失血性休克的危重疾病。由于肺部特别 容易受到创伤继发感染的影响，并且也容易受到中性粒细胞介导的损伤，该模型 提供了一种评估新目标/策略的方法，以促进中性粒细胞功能的选择性方面 增强宿主防御而不加剧组织损伤。最后，我们将应用人体创伤患者 材料（血液、血浆、肺液）到这些方法中以探测疾病特异性机制 中性粒细胞反应。由于我们的研究将探讨基本的生物机制，例如整合素 激活后，这项工作也将在多个领域产生广泛的影响。这些综合方法共同将 通过跨越分子、细胞和生物体/疾病来解决中性粒细胞生物学的主要知识空白 实验水平。