Applying Innovative Lung Mapping Strategies to Understand Alveolar Capillary Barrier Permeability in ARDS

项目摘要 此R35应用程序描述了一个强大的研究框架，可帮助发现和翻译新的 急性呼吸窘迫综合征(ARDS)的治疗靶点 失败会导致高死亡率，并且没有有益的靶向治疗。ARDS仍然是一个重要的 每年影响19万美国人的健康问题，花费数十亿美元，并留下大部分 患者死亡或严重受损。需要对发病机制有新的见解来深化我们的 了解导致ARDS的潜在机制，以及开发新的治疗方法。 因此，确定可研究的临床相关治疗靶点的需求尚未得到满足。 机械地、快速地进行了强有力的临床前研究。在过去的14年里，我一直是 让我的研究团队走在翻译发现的前列。我的团队已经取得了重大进展 对了解ARDS病理生理学的贡献。我的实验室的主要重点是确定关键 ARDS基础肺泡毛细血管屏障功能和功能障碍的细胞和分子调节 病理学。我的R35研究计划旨在利用肺组织识别ARDS中的新介质 单细胞水平上基因表达谱的成像质谱学和深度表型分析 用先进的统计方法识别领先目标。将在体外转基因中研究新的靶点 定义细胞和分子机制的模型系统，以促进新技术的开发 治疗药物将在临床前模型中进行测试。我的研究框架围绕三个目标展开：发现、 机制与翻译。目标1-发现。为了实现这一目标，我们将使用 成像质谱学和单细胞RNA测序以创建表达谱和蛋白质“图谱” 受伤的和未受伤的人的肺。使用先进的统计方法，我们将确定有希望的 推进进一步研究的目标。目标2--机制。利用我们现有的肺损伤模型， 制度资源和新方法，我们将定义导致 通过建立新的转基因细胞和小鼠株系来治疗ARDS的肺泡毛细血管屏障通透性 机械学研究。目标3--翻译。建立在我们现有的体外人肺和体内小鼠的基础上 模型，我们将根据我们的Discovery和 机理研究。有了这个R35，我的实验室将通过以下方式推进NHLBI的任务：生成 急性损伤和未损伤人的单细胞转录谱和组织蛋白表达水平 肺，确定ARDS的新治疗靶点，确定细胞和分子调控机制 在小鼠和人类模型上进行肺泡毛细血管屏障功能障碍和临床前研究。R35 将为我在ARDS领域开辟新天地提供必要的支持和灵活性。

Project Summary This R35 application describes a robust research framework to facilitate discovery and translation of new therapeutic targets in Acute Respiratory Distress Syndrome (ARDS), a common cause of acute respiratory failure that carries a high mortality rate and has no beneficial targeted therapies. ARDS remains a significant health problem affecting 190,000 Americans per year, costing billions of dollars, and leaving the majority of patients dead or significantly impaired. New insights into the pathogenesis are needed to deepen our understanding of the underlying mechanisms that lead to ARDS as well as to develop novel therapeutics. Thus, there is an unmet need to define clinically relevant therapeutic targets that can be studied mechanistically and rapidly carried through robust pre-clinical studies. For the last 14 years I have been building my research team to be on the forefront of translational discovery. My group has made major contributions to understanding ARDS pathophysiology. The major focus of my laboratory is defining the key cellular and molecular regulators of alveolar capillary barrier function and dysfunction that underlies ARDS pathology. My R35 research program is designed to identify novel mediators in ARDS using lung tissue imaging mass spectrometry and deep phenotyping of gene expression profiles at the single cell level coupled with advanced statistical methods to identify leading targets. New targets will be studied in in vitro transgenic model systems to define cellular and molecular mechanisms in order to facilitate the development of novel therapeutics to be tested in pre-clinical models. My research framework is centered on three goals: Discovery, Mechanism and Translation. Goal 1 – Discovery. To accomplish this goal, we will break new ground using imaging mass spectrometry and single cell RNA sequencing to create an expression profile and protein “map” of the injured and uninjured human lung. Using advanced statistical approaches, we will identify promising targets to take forward into further studies. Goal 2 – Mechanism. Leveraging our existing lung injury models, institutional resources and new approaches, we will define the fundamental pathologic mechanisms that lead to alveolar capillary barrier permeability in ARDS by generating novel transgenic cell and mouse lines for mechanistic studies. Goal 3 – Translation. Building on our existing ex vivo human lung and in vivo mouse models, we will generate rigorous pre-clinical data based on new targets identified in our Discovery and Mechanism studies. With this R35, my lab will advance the mission of the NHLBI by: generating a catalogue of single cell transcription profiles and tissue protein expression levels in acutely injured and uninjured human lung, identifying novel therapeutic targets in ARDS, defining the cellular and molecular mechanisms regulating alveolar capillary barrier dysfunction and conducting pre-clinical studies in mouse and human models. The R35 will provide the support and flexibility necessary for me to break new ground in ARDS.