Cytoskeletal Regulation of Lung Endothelial Pathobiology in ARDS

摘要： 肺内皮细胞(ECs)和ARDS血管内型在ARDS病理生物学中的关键作用 在当前的全球新冠肺炎大流行中被戏剧性地凸显出来。这一高度翻译的PPG续订 将讨论未加控制的EC通透性对毁灭性的多器官衰竭和死亡率的贡献 ARDS通过在分子和基因组水平上对血管屏障调节基因的全面了解来实现。 整形和修复。这次更新保持了对EC细胞骨架在病理生物学中的关键作用的关注 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- 批准的ARDS药物疗法，可减轻肺血管通透性和炎症。我们建议 4个与临床高度相关的紧密编织的PPG项目，以特定的肺EC靶蛋白/基因为中心 参与：i)ARDS中未加控制的血管通透性和损伤；ii)血管对刺激的反应- Vili的严重机械应力；三)导致非洲ARDS健康差异的遗传基础 以及(Iv)提供新的ARDS治疗机会。所有领域的主题整合 项目包括PPG靶基因中与ARDS相关的SNPs和CpG位点的功能表征以及 ROS在外周细胞骨架重塑的转录和生化调节中的作用 细胞骨架驱动的片状脂溢症和局部黏附(FA)重组，促进EC缝隙闭合。项目 #1将利用系统生物学方法定义EC屏障反应的新的细胞骨架调节 多功能非肌肉肌球蛋白轻链激酶亚型(NmMLCK)及其细胞骨架结合伙伴， Cortactin。项目2(一种新的PPG添加)扩展了我们最近的报告，即EC分泌的损害相关 分子模式(DAMP)蛋白eNAMPT(烟酰胺磷酸核糖转移酶)在急性呼吸窘迫综合征中起关键作用。 炎性通透性和损伤通过eNAMPT结扎Toll样受体4(TLR4)。项目3将与 整合素β4和Kindlin2对细胞间双向信号转导的调节 骨骼和局部粘连(FA)动力学，促进板脂介导的炎症闭合。 引入EC缺口。项目4(一个新的PPG项目)将阐明关键障碍-监管重新调整之间的相互作用 受体：调节白细胞的鞘氨醇-1-磷酸(S1P)受体1(S1PR1)、S1PR3和P-选择素 P-选择素糖蛋白配体1(PSGL1)结扎肺血管再通和肺血管漏。由四个人支持 高度互动的核心编织到每个PPG项目中(管理、基因组/蛋白质组、临床前动物 模型，生物物理成像)，这个PPG将继续利用出色的科学环境 亚利桑那大学和伊利诺伊大学，以及Interactive Basic优秀的PPG翻译团队 以及内科科学家调查人员。除了告知ARDS健康差异的遗传基础外，我们的 这项工作直接解决了对肺内皮细胞屏障调节机制的更深入了解的迫切需求。 以及恢复受损肺循环完整性以降低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.