Pre-Clinical Large and Small Animal Models of ARDS/VILI

摘要： 急性肺损伤临床前模型核心(Core C)旨在为PPG研究人员提供 严格定义且可复制的组合性急性呼吸窘迫综合征大鼠和猪模型 (ARDS)和呼吸机所致肺损伤(VILI)。史无前例的高死亡率新冠肺炎大流行 新冠肺炎引发的急性呼吸窘迫综合征的发病率极大地提高了对更深入地了解严重急性呼吸窘迫综合征的需求 EC细胞骨架在ARDS和VILI病理生物学中的作用并加深了FDA未得到满足的需求- 批准的急性呼吸窘迫综合征药物疗法。核心C将全面生成、管理和提供所有动物- 通过实现五个具体目标，为所有四个项目提供相关的实验、资源和专业知识。特定的 AIM#1提供了一整套的专业知识、培训、设备和数据分析工具来进行广泛的研究 并描述了细胞骨架在肺损伤的临床前模型中的作用。核心C将采用状态- A)表征细胞骨架在调节肺内皮细胞(EC)屏障中的作用 职能，b)确定具体干预措施的效果，以便深入了解 新的治疗策略，以及c)促进将基础研究转化为临床干预措施。特定的 目标2是饲养和维护在这个PPG中使用的老鼠和猪。具体目标#3有选择性地审查 药物激动剂、拮抗剂或针对PPG靶向效应器、细胞骨架和 粘着斑蛋白作为ARDS/VILI模型潜在的治疗策略和途径。特定目标 #4提供严格执行、协议驱动的特定实验策略性能，包括 ARDS/VILI的临床前模型。具体目标5是执行高度详细的研究并提供高质量 基因组、生化表型(BAL蛋白、BAL细胞计数/细胞数、肺组织白蛋白水平、 肺毛细血管渗漏的伊文思蓝染色(EBD)，肺组织学定量，和 免疫组织化学)。此外，核心C还提供生理测量(静态和动态顺应性 呼吸系统、氧合指数)、放射照片和超声图像支持这些研究。这个 ARDS的大小和恢复反应将通过产生急性肺损伤的严重程度来确定 得分(ALISS)。核心C将进行肺损伤评估，向个人提供血浆和组织样本 用于特定检测的项目(包括免疫组织化学和蛋白质印迹分析)。我们在核心C中的工作将 为所有四个项目提供全面的临床前支持，并提供他们需要的工具以获得更大的机械性 理解肺内皮细胞屏障调节，同时推动治疗进展，以恢复 改善受损肺循环的完整性，从而降低ARDS死亡率。

ABSTRACT: The Pre-Clinical models of acute lung injury Core (Core C) is designed to provide PPG investigators with rigorously defined and reproducible rat and porcine models of combined acute respiratory distress syndrome (ARDS) and ventilation-induced lung injury (VILI). The unprecedented COVID-19 pandemic with high mortality rates of COVID-19-induced ARDS has dramatically raised the demand for a deeper understanding of the critical role of the EC cytoskeleton in the pathobiology of ARDS and VILI and deepened the unmet need for FDA- approved ARDS pharmacotherapies. Core C will comprehensively generate, manage, and provide all animal- related experiments, resources, and expertise to all four projects by accomplishing five specific aims. Specific Aim #1 provides a complete range of expertise, training, equipment, and data analysis tools to extensively study and characterize the role of the cytoskeleton in preclinical models of lung injury. Core C will employ the state-of- the-art techniques to a) characterize the role of the cytoskeleton in regulating lung endothelial cell (EC) barrier function, b) determine the effects of specific interventions to provide insight into the efficacy and mechanisms of novel therapeutic strategies, and c) facilitate the translation of basic research to clinical interventions. Specific Aim #2 is to house and maintain rats and pigs utilized in this PPG. Specific Aim #3 examines selective pharmacological agonists, antagonists, or monoclonal antibodies for PPG-targeted effectors, cytoskeletal, and focal adhesion proteins as potential therapeutic strategies and approaches for ARDS/VILI models. Specific Aim #4 provides rigorously performed, protocol-driven performance of specific experimental strategies involving preclinical models of ARDS/VILI. Specific Aim #5 is to perform highly detailed studies and provide high quality genomic, biochemical phenotyping (BAL protein, BAL cell count/cellularity, lung tissue albumin levels, assessment of lung capillary leakage by Evans Blue dye (EBD), quantitative lung histology, and immunohistochemistry). Also, Core C provides physiologic measurements (static and dynamic compliance of the respiratory system, oxygenation index), radiographic and ultrasound images to support these studies. The magnitude of ARDS and recovery responses will be determined by generating the acute lung injury severity score (ALISS). Core C will perform lung injury assessment, provide plasma, and tissue samples to individual projects for specific assays (including immunohistochemistry and western blot analysis). Our work in Core C will provide full preclinical support to all four projects and provide the tools they need to get a greater mechanistic understanding of lung EC barrier regulation while driving therapeutic developments directed toward restoring the integrity of the injured pulmonary circulation, thereby reducing ARDS mortality.