Lung epithelial cell-derived C3 in acute lung injury

肺上皮细胞衍生的 C3 在急性肺损伤中的作用

基本信息

批准号：
10720687
负责人：
Hrishikesh Satish Kulkarni
金额：
$ 59.85万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2027-09-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10720687
关键词：
Acute Lung Injury Acute Respiratory Distress Syndrome Address Affect Alveolar Alveolar Macrophages Area Binding Bioinformatics Biological Assay Biology CRISPR screen Cell Compartmentation Cell Death Cell Death Induction Cell Survival Cell physiology Cells Cessation of life Characteristics Circulation Clustered Regularly Interspaced Short Palindromic Repeats Compensation Complement Complement 3a Complement 3b Complement Factor B Cytoprotection Data Dependence Epithelial Cells Epithelium Experimental Models Extracellular Space Gene Delivery Genetic Goals Host Defense Host Defense Mechanism Human Immune Immune response In Vitro Inflammatory Injury Knock-out Knockout Mice Knowledge Libraries Liver Lung Macrophage Mediating Modeling Morbidity - disease rate Mus Myeloid Cells National Heart, Lung, and Blood Institute Phagocytosis Population Predisposition Principal Investigator Process Production Proteins Pseudomonas aeruginosa Pseudomonas aeruginosa pneumonia Publishing Research Personnel Research Priority Role Site Source Stress Testing Thinking Time Tissues Transgenic Mice Wild Type Mouse Work airway epithelium alveolar epithelium cell injury cell type complement C3 precursor complement system conditional knockout cytokine design disability extracellular human model immune cell infiltrate improved in vivo in vivo Model lung injury mortality mouse model novel therapeutics overexpression pathogen protective effect receptor resilience response response to injury uptake

项目摘要

PROJECT SUMMARY/ABSTRACT The goal of this R01 proposal is to investigate how complement component C3 derived from the lung epithelium can improve host resilience during acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). C3 is a central component of the complement system and one of the most abundant circulatory proteins. It is an early responder to injury and operates by killing pathogens and clearing debris. However, we have shown that it takes time for circulating C3 to reach the alveolar space in ALI, thereby increasing the dependence on C3 present in the lung. We have also shown that not only is lung epithelial cell-derived C3 secreted, but also its intracellular stores are central to the protection against ALI and cell death. These findings are a shift from the conventional thinking that liver-derived C3 is the main operational form of complement in the lung and increases the emphasis on epithelial cells as a source. Additionally, we have shown that many immune cell types—including macrophages—derive C3 stores via uptake from the extracellular space, and this uptake improves their effector response. Hence, this proposal focuses on an emerging role for lung epithelial cell-derived C3 in promoting the survival and function of epithelial cells and macrophages, each one vital in ALI. A major hurdle for investigating tissue-specific roles of complement has been the limited availability of models and assays. We have developed new transgenic mouse models and functional assays that distinguish the roles of liver- and lung-derived complement and show that epithelial cell-derived C3 is central to protection in ALI. We have supplemented these models and assays with data from ex vivo human models to show key roles for C3 in the cellular response to injury. These results support our central hypothesis that lung epithelial cell-derived C3 modulates the bronchoalveolar epithelial and immune cell niche to reduce ALI. This proposal will test our hypothesis by achieving two Specific Aims. Aim 1 compares targeted liver derived C3-deficient mice with lung epithelial cell- derived C3-deficient mice to assess if the cytoprotection offered by lung epithelial cell-derived C3 is cell-type intrinsic or also regional. It will also address how to optimally augment C3 to protect the injured lung. Aim 2 uses a combination of conditional knockout mice and deceased human donor lungs to assess how lung epithelial cell- derived C3 influences the pulmonary myeloid cell compartment, especially alveolar macrophage survival and function. It also addresses how to modulate C3 uptake in these cells to optimize their function. The proposal brings together a principal investigator with expertise in complement biology and acute lung injury, and co-investigators with expertise in gene delivery, CRISPR screens and bioinformatics to determine the immunobiological role of lung epithelial cell-derived C3 in facilitating host defense at the site of injury and promoting tissue resilience. The proposed work is important because understanding how the early host immune response modulates tissue damage is essential for designing and implementing new therapies for ALI. The knowledge will form the basis of locally delivered, host-focused therapies for ARDS, thus aligning with a priority research area for the NHLBI.

项目概要/摘要该 R01 提案的目标是研究补体成分 C3 如何源自肺上皮可以提高急性肺损伤（ALI）和急性呼吸窘迫综合征（ARDS）期间宿主的恢复能力。是补体系统的核心组成部分，也是最丰富的循环蛋白之一。对损伤做出早期反应并通过杀死病原体和清除碎片来进行操作。但是，我们已经证明它是这样的。在 ALI 中，循环 C3 需要时间才能到达肺泡腔，从而增加了对存在的 C3 的依赖性我们还发现，不仅肺上皮细胞分泌 C3，而且其细胞内也分泌 C3。这些发现是对传统疗法的转变。认为肝源性 C3 是肺中补体的主要运作形式增加了重点此外，我们还发现许多免疫细胞类型，包括上皮细胞。巨噬细胞——通过从细胞外空间摄取来衍生 C3 储存，这种摄取改善了它们的效应器因此，本提案重点关注肺上皮细胞来源的 C3 在促进反应中的新兴作用。上皮细胞和巨噬细胞的存活和功能，这两者在 ALI 中都至关重要，这是研究的主要障碍。补体的组织特异性作用一直是我们开发的模型和检测方法有限的原因。新的转基因小鼠模型和功能测定可区分肝源性和肺源性的作用补充并表明上皮细胞来源的 C3 对于 ALI 的保护至关重要。使用离体人体模型的数据进行模型和检测，以显示 C3 在细胞反应中的关键作用这些结果支持我们的中心假设，即肺上皮细胞来源的 C3 调节损伤。支气管肺泡上皮和免疫细胞生态位以减少 ALI 该提案将通过以下方式检验我们的假设。实现两个具体目标 1 将目标肝脏来源的 C3 缺陷小鼠与肺上皮细胞进行比较。衍生的 C3 缺陷小鼠，以评估肺上皮细胞衍生的 C3 提供的细胞保护是否是细胞类型它还将讨论如何最佳地增强 C3 以保护受伤的肺部。条件性基因敲除小鼠和已故人类供体肺的组合，以评估肺上皮细胞如何衍生的 C3 影响肺髓细胞区室，特别是肺泡巨噬细胞的存活和该提案还解决了如何调节这些细胞中的 C3 摄取以优化其功能。具有补体生物学和急性肺损伤专业知识的首席研究员和共同研究员拥有基因传递、CRISPR 筛选和生物信息学方面的专业知识，以确定免疫生物学作用肺上皮细胞衍生的 C3 可促进损伤部位的宿主防御并促进组织弹性。拟议的工作很重要，因为了解早期宿主免疫反应如何调节组织损伤对于设计和实施 ALI 的新疗法至关重要。这些知识将构成治疗 ALI 的基础。本地提供的针对 ARDS 的以宿主为中心的疗法，从而与 NHLBI 的优先研究领域保持一致。