Selective modulation of neutrophils in critical illness

摘要 危重疾病描述由可能发生的免疫功能障碍引起的危及生命的状态。 在创伤、休克、烧伤或大手术后。创伤诱导的免疫功能低下增强 对继发感染的易感性，这是医院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，对宿主抵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，它们也会导致旁观者组织损伤 阿森纳。创伤对中性粒细胞的影响导致免疫功能障碍，包括明显的缺陷。 发现和抵抗感染的中性粒细胞能力，以及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 招募和效应器在健康和疾病中的作用。这个Mira应用程序描述了我们的协同 危重病期间中性粒细胞病理生物学机制的识别和理解方法。 首先，我们建立了一种创新的方法，在体外获得中性粒细胞，绕过了技术上的 他们的基因改造的障碍。这使我们能够执行多路正向基因筛查 CRISPR-Cas9在中性粒细胞中，从而确定其招募的新机制方面(例如，整合素 激活)和抗菌功能(例如，吞噬)。第二，我们已经建立了一个小鼠模型。 继发于失血性休克的呼吸道感染所致的危重疾病。因为肺特别是 此模型易受创伤继发感染，也易受中性粒细胞介导的损伤 提供一种方法来评估新的目标/策略，以促进中性粒细胞功能的选择性方面 在不加剧组织损伤的情况下增强宿主防御。最后，我们将应用人类创伤患者 材料(血液，血浆，肺液)这些途径，以探讨疾病的特异性机制 中性粒细胞反应。由于我们的研究将询问基本的生物学机制，如整合素 激活后，这项工作还将对多个领域产生广泛影响。综合起来，这些综合方法将 通过跨越分子、细胞和生物/疾病来解决中性粒细胞生物学方面的主要知识差距 实验的水平。

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.