GasderminD regulation of Acute Lung Injury

摘要： 肺内皮细胞屏障功能丧失和内皮细胞(EC)死亡是主要的致病特征 急性呼吸窘迫综合征(ARDS)。尽管看起来基本的免疫细胞过程 是ARDS介导的血管损伤和功能障碍的核心，也是内皮细胞的实际机制 由免疫细胞调控的死亡在很大程度上仍是未知的。因此，目前还没有有效的 药物疗法。因此，为了开发更有效的治疗策略，有必要 了解免疫细胞功能与血管功能障碍之间的确切联系机制。 这一建议侧重于一种新的分子范例，即免疫细胞(单核细胞)驱动内皮细胞 细胞死亡。我们广泛的支持性数据使我们形成了与肺有关的总体假设 ARDS时血管内皮细胞损伤对单核/巨噬细胞来源的GasderminD嗜热功能的影响 (GsdmD)被包裹在循环微粒(MP)中，与嘌呤能受体P2X7结合。至 为了确定这一新途径的特异性，我们提出了以下具体目标。在Aim1，我们将 单核细胞来源的GasderminD在介导内皮细胞下垂机制中的作用 阿里。我们将利用现有的小鼠模型以及ARDS患者的体外和体外模型来研究 GsdmD介导内皮损伤。在AIM2中，我们将确定MP封装的GsdmD的调节 单核细胞的细胞病变功能。我们将研究GasderminD的磷酸化调节机制 它的细胞病变行为和对泛素介导的降解的脆弱性。最后，在Aim3中，我们将描绘 P2X7在MP-GsdmD介导的内皮细胞死亡机制中的作用我们会研究 功能性细胞外MP相关的GsdmD与靶细胞结合的机制 嘌呤能受体P2X7。 这些研究将首次阐明细胞外GsdmD的损伤行为与 嘌呤能受体P2X7被包裹在MPS中，可能在肺损伤中起中心作用。签立 这些研究将为细胞相互作用的重大机制进展奠定基础 免疫细胞和内皮之间的关系，这可能为设计新的治疗靶点提供机会 减轻肺血管损伤的严重程度。

Abstract: The loss of lung endothelial barrier function and endothelial cell (EC) death is a primary pathogenic feature of acute respiratory distress syndrome (ARDS). Although it appears that fundamental immune cell processes are at the core of ARDS-mediated vascular injury and dysfunction, the actual mechanism of endothelial cell death regulated by immune cells remains largely unknown. As a result there are currently no effective pharmacologic therapies. In order to develop more effective therapeutic strategies, it is therefore necessary to understand the precise mechanisms that link immune cell function to vascular dysfunction. This proposal focuses on a new molecular paradigm whereby immune cells (monocytes) drive endothelial cell death. Our extensive supportive data have lead us to formulate the overarching hypothesis that links lung endothelial injury during ARDS to the pyroptotic function of monocyte/macrophage derived GasderminD (GsdmD) encapsulated in circulating microparticles(MP) in association with purinergic receptor, P2X7. To determine the specifics of this novel pathway, we propose the following specific aims. In Aim1, we will delineate the role of monocyte derived GasderminD in mediating endothelial pyroptosis in the mechanism of ALI. We will use available murine model along with in vitro and ex vivo model of ARDS patients to study GsdmD mediated endothelial injury. In Aim2, we will determine the regulation of MP encapsulated GsdmD’s cytopathic function by monocytes. We will study the mechanism how phosphorylation of GasderminD regulates its cytopathic behavior and vulnerability to ubiquitin-mediated degradation. Finally, in Aim3, we will delineate the role of P2X7 in the mechanism of MP GsdmD mediated endothelial cell death. We will study the mechanisms of extracellular MP-associated GsdmD engagement to target cells mediated by the functional purinergic receptor P2X7. These studies will be the first to elucidate the injurious behavior of extracellular GsdmD in coordination with the purinergic receptor, P2X7 encapsulated in MPs, which may play a central role in lung injury. Execution of these studies will lay the foundation for a significant mechanistic advance regarding the cellular interplay between immune cells and endothelia that may provide opportunities for devising novel therapeutic targets that lessen the severity of lung vascular injury.