Role of intravascular ERO1@ in acute lung injury

项目摘要 急性肺损伤(ALI)是一种危及生命的疾病，在美国每年影响20万名患者。 与肺炎、败血症和创伤有关，导致肺功能不全，最终导致多系统 器官衰竭。由于激活的中性粒细胞过度募集到肺微血管是导致 ALI，更好地理解中性粒细胞-内皮细胞相互作用的机制将提供 洞察开发一种治疗ALI的有效疗法。我们之前证明了血管内 蛋白二硫键异构酶(PDI)增强中性粒细胞和血小板表面受体的配体结合活性 并在血管炎症期间导致血管内细胞与细胞之间的相互作用。为了阐明细胞外PDI是如何 我们发现，活动是受调控的，调控机制是否参与了ALI的病理 内质网氧化还原蛋白1α(ERO 1α，内质网氧化还原蛋白1的关键酶)促进中性粒细胞的生长 在血管炎症期间重新招募。在这项提议中，我们将检验血管内ERO 1α的假设 通过修饰二硫键增强中性粒细胞黏附受体的配体结合功能，诱导 中性粒细胞重新聚集到急性肺部炎症部位。在目标1中，我们将确定 ERO 1α调控中性粒细胞与内皮细胞的相互作用。在目标2中，我们将利用肺部活体成像技术来 探讨血管内ERO_1α在ALI中的病理作用。在目标3中，使用急性白血病患者的血液 呼吸窘迫综合征(ARDS)，我们将确定ERO 1α在疾病进展中的作用 以及ARDS患者的严重程度。这些研究将利用生化、细胞生物学、遗传学和共聚焦技术。 活体成像方法。由于对胞外硫醇修饰机制知之甚少，建议的 研究将确定促进ALI/ARDS发病机制的基本机制，但尚未探索。

Project Summary Acute lung injury (ALI) is a life-threatening condition, which affects > 200,000 patients annually in the U.S. It is associated with pneumonia, sepsis, and trauma, leading to pulmonary insufficiency and eventually multisystem organ failure. Since excessive recruitment of activated neutrophils to lung microvessels is a primary cause of ALI, a better understanding of the mechanism mediating neutrophil-endothelial cell interactions will provide insight into developing an effective therapeutic for treating ALI. We previously demonstrated that intravascular protein disulfide isomerase (PDI) enhances the ligand-binding activity of neutrophil and platelet surface receptors and leads to intravascular cell-cell interactions during vascular inflammation. To elucidate how extracellular PDI activity is regulated and whether the regulatory mechanism contributes to the pathology of ALI, we have found that endoplasmic reticulum oxidoreductin 1α (ERO1α, a key oxidase of PDI in the ER) promotes neutrophil recruitment during vascular inflammation. In this proposal, we will test the hypothesis that intravascular ERO1α enhances the ligand-binding function of neutrophil adhesion receptors by modifying disulfide bonds, inducing neutrophil recruitment to sites of acute lung inflammation. In Aim 1, we will determine the mechanism by which ERO1α regulates neutrophil-endothelial cell interactions. In Aim 2, we will utilize lung live imaging techniques to investigate the pathological role of intravascular ERO1α in ALI. In Aim 3, using the blood of patients with acute respiratory distress syndrome (ARDS), we will determine the contribution of ERO1α to the disease progression and severity in patients with ARDS. These studies will employ biochemical, cell biological, genetic, and confocal intravital imaging approaches. Since little is known about extracellular thiol-modifying machinery, the proposed studies will identify an essential, yet unexplored, mechanism that promotes the pathogenesis of ALI/ARDS.