Cytoskeletal Regulation of Lung Endothelial Pathobiology in ARDS

摘要： 肺内皮细胞（EC）和ARDS血管内皮型在ARDS病理学中的关键作用是 在当前的全球COVID-19大流行中，这一点得到了戏剧性的强调。这种高度转化的PPG更新 将解决未受抑制的EC渗透性对破坏性多器官衰竭和死亡率的贡献， 通过在分子和基因组水平上全面了解血管屏障调节因子， 测量和修复。这一更新保持其重点的关键作用EC细胞骨架的病理生物学 ARDS和呼吸机诱导的肺损伤（VILI），并寻求直接解决FDA未满足的需求- 已批准的减轻肺血管通透性和炎症的ARDS药物疗法。我们提出 4个高度临床相关、紧密结合的PPG项目集中于特定的肺EC靶蛋白/基因， 参与：i）ARDS中未受抑制的血管通透性和损伤; ii）血管对过量- VILI中的机械压力; iii）促进非洲人ARDS健康差异的遗传基础 血统受试者;和，iv）提供新的ARDS治疗机会。所有主题的整合 项目包括ARDS相关SNP和PPG靶基因中CpG位点的功能表征， ROS在外周细胞骨架重塑、 胞浆驱动的板状伪足和促进EC间隙闭合的粘着斑（FA）重组。项目 #1将利用系统生物学方法来定义EC屏障反应的新细胞骨架调节， 多功能非肌肉肌球蛋白轻链激酶同种型（nmMLCK）及其细胞内结合配偶体， corn。项目#2（新的PPG添加）扩展了我们最近的报告，即EC分泌的损伤相关的 分子模式（DAMP）蛋白，eNAMPT（烟酰胺磷酸核糖基转移酶），是ARDS的关键， 通过Toll样受体4（TLR 4）的eNAMPT连接的炎性渗透性和损伤。项目#3将在- 通过整合素β4（ITGB 4）和kindlin 2对细胞与神经元之间的双向信号传导进行新的调节， 骨骼和局灶性粘连（FA）动力学，促进板层脂醛介导的炎症关闭， 导致EC差距。项目#4（一个新的PPG项目）将阐明关键障碍-监管重新 受体：1-磷酸鞘氨醇（S1 P）受体1（S1 PR 1）、S1 PR 3和P-选择素，调节白细胞 通过P-选择素糖蛋白配体1（PSGL 1）的结扎引起的募集和肺血管渗漏。支持四 每个PPG项目（行政、基因组/蛋白质组、临床前动物）都包含高度互动的核心 模型，生物物理成像），该PPG将继续利用优秀的科学环境， 亚利桑那大学和伊利诺伊大学，以及优秀的PPG互动基础翻译团队 和医学科学家调查员。除了告知ARDS健康差异的遗传基础外，我们的 工作直接解决了迫切的未满足的需要，更大的机制了解肺EC屏障调节， 恢复受损肺循环的完整性以降低ARDS死亡率的新疗法。

ABSTRACT: The critical role of lung endothelial cells (ECs) and the ARDS vascular endotype in ARDS pathobiology is being dramatically highlighted in the current global COVID-19 pandemic. This highly translational PPG renewal will address the contribution of unchecked EC permeability to the devastating multi-organ failure and mortality of ARDS by providing a comprehensive understanding at the molecular and genomic level of vascular barrier reg- ulation and repair. This renewal maintains its focus on the critical role of the EC cytoskeleton in the pathobiology of ARDS and ventilator-induced lung injury (VILI) and seeks to directly address the unmet need for FDA- approved ARDS pharmacotherapies that attenuate lung vascular permeability and inflammation. We propose 4 highly clinically-relevant, tightly-woven PPG Projects centered on specific lung EC target proteins/genes that are involved in: i) the unchecked vascular permeability and injury in ARDS; ii) vascular responses to exces- sive mechanical stress in VILI; iii) contributing to the genetic basis for ARDS health disparities in African descent subjects; and, iv) providing novel ARDS therapeutic opportunities. Thematic integration across all projects includes functional characterization of ARDS-associated SNPs and CpG sites in PPG target genes and the role of ROS in transcriptional and biochemical regulation of peripheral cytoskeletal remodeling, formation of cytoskeletal-driven lamellipodia and focal adhesion (FA) reorganization that promotes EC gap closure. Project #1 will utilize system biology approaches to define novel cytoskeletal regulation of EC barrier responses by the multi-functional non-muscle myosin light chain kinase isoform (nmMLCK) and its cytoskeletal-binding partner, cortactin. Project #2 (a new PPG addition) extends our recent report that EC secretion of the damage-associated molecular pattern (DAMP) protein, eNAMPT (nicotinamide phosphoribosyltransferase), is critical to ARDS in- flammatory permeability and injury via eNAMPT ligation of the Toll-like receptor 4 (TLR4). Project #3 will inter- rogate the novel regulation by integrin β4 (ITGB4) and kindlin2, of the bidirectional signaling between the cyto- skeleton and focal adhesion (FA) dynamics which promotes lamellipodial-mediated closure of inflammation-in- duced EC gaps. Project #4 (a new PPG Project) will elucidate interactions between key barrier-regulatory re- ceptors: sphingosine-1-phosphate (S1P) receptor 1 (S1PR1), S1PR3, and P-selectin which regulate leukocyte recruitment and lung vascular leak via ligation by P-selectin glycoprotein ligand 1 (PSGL1). Supported by four highly interactive cores woven into each PPG Project (Administrative, Genome/Proteome, Preclinical Animal Model, Biophysical Imaging), this PPG will continue to leverage the outstanding scientific environments at the University of Arizona and University of Illinois, and the outstanding PPG translational team of interactive basic and physician-scientist investigators. In addition to informing the genetic basis for ARDS health disparities, our work directly addresses the urgent unmet needs for greater mechanistic understanding of lung EC barrier regu- lation and novel therapeutics that restore the integrity of the injured lung circulation to reduce ARDS mortality.