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

摘要： 急性肺损伤核心临床前模型（核心C）旨在为PPG研究者提供 急性呼吸窘迫综合征的严格定义和可重复的大鼠和猪模型 （ARDS）和通气诱导的肺损伤（VILI）。史无前例的高死亡率COVID-19大流行 COVID-19引起的ARDS的发病率大大提高了对更深入了解关键因素的需求。 EC细胞骨架在ARDS和VILI病理生物学中的作用，并加深了FDA未满足的需求， 批准的ARDS药物治疗。核心C将全面产生，管理，并提供所有动物- 通过实现五个具体目标，将实验、资源和专业知识与所有四个项目相关联。具体 目标#1提供一整套专业知识、培训、设备和数据分析工具， 并表征细胞骨架在肺损伤临床前模型中的作用。核心C将采用国家- 现有技术a）表征细胞骨架在调节肺内皮细胞（EC）屏障中的作用 功能，B）确定具体干预措施的效果，以深入了解 新的治疗策略，以及c）促进基础研究向临床干预的转化。具体 目的#2是圈养和饲养本PPG中使用的大鼠和猪。具体目标#3检查选择性 PPG靶向效应物的药理学激动剂、拮抗剂或单克隆抗体，细胞骨架， 粘着斑蛋白作为ARDS/VILI模型的潜在治疗策略和方法。具体目标 #4提供了严格执行的，方案驱动的特定实验策略的性能， ARDS/VILI的临床前模型。具体目标#5是执行非常详细的研究，并提供高质量的 基因组、生化表型（BAL蛋白、BAL细胞计数/细胞构成、肺组织白蛋白水平， 通过伊文思蓝染料（EBD）评估肺毛细血管渗漏，定量肺组织学， 免疫组织化学）。此外，核心C提供生理测量（静态和动态顺应性）， 呼吸系统、氧合指数）、放射学和超声图像来支持这些研究。的 通过生成急性肺损伤严重程度来确定ARDS的程度和恢复反应 评分（ALISS）。核心C将进行肺损伤评估，向个体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Core C中的工作将 为所有四个项目提供全面的临床前支持，并提供他们所需的工具，以获得更大的机制， 了解肺EC屏障调节，同时推动治疗发展， 损伤的肺循环的完整性，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.