Cytoskeletal Regulation of Lung Endothelial Pathobiology

肺内皮病理学的细胞骨架调节

• 批准号: 9925241
• 负责人: Joe G. N. Garcia
• 金额: $ 233.57万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9925241
• 关键词: ABL1 gene; Actin-Binding Protein; Actins; Adult Respiratory Distress Syndrome; Affect; African; Agonist; Antibodies; Arizona; Attenuated; Biophysics; Blood Vessels; Breathing; Cells; Collaborations; Cytoskeleton; Development; Disease; Dissociation; EMS1 gene; Endothelial Cells; Endothelium; Environment; Event; F-Actin; Focal Adhesion Kinase 1; Focal Adhesions; Genetic; Illinois; Image; In Vitro; Individual; Inflammation; Integrin beta Chains; Integrin beta4; Intercellular Junctions; Knowledge; Lung; MYLK gene; Mechanical Stress; Mechanical ventilation; Mediating; Membrane; Molecular; Myosin Light Chain Kinase; Outcome; Participant; Pathway interactions; Patients; Periodicity; Peripheral; Permeability; Phase; Physicians; Population; Post-Translational Protein Processing; Pre-Clinical Model; Production; Protein Isoforms; Proteins; Pulmonary Circulation; Regulation; Regulator Genes; Research Personnel; Resolution; Role; Scientist; Signal Transduction; Single Nucleotide Polymorphism; Stains; Stimulus; Stress Fibers; Stretching; Structure; TNF gene; Testing; Thrombin; Universities; Variant; Vascular Endothelial Growth Factors; Vascular Permeabilities; Ventilator-induced lung injury; biophysical analysis; clinically relevant; genetic analysis; health disparity; high risk; inhibitor/antagonist; injured; mortality; new therapeutic target; non-muscle myosin; novel; paxillin; polymerization; protein complex; response; restoration; therapeutic development; therapeutic target; therapy design; translational model; vasodilator-stimulated phosphoprotein

DESCRIPTION (Provided by applicant): This revised A1 PPG application remains a highly translational PPG focused on the critical role of the endothelial cell (EC) cytoskeleton in the pathobiology of acute respiratory distress syndrome (ARDS) and ventilator-induced lung injury (VILI). ARDS, a devastating disorder with a mortality of ~30-35%, is defined by increases in lung vascular permeability, a major influence on ARDS mortality. The clinically-relevant PPG studies we have proposed will define the molecular mechanisms by which EC signaling drives cytoskeletal remodeling to either disrupt vascular integrity via production of paracellular gaps (that increases lung vascular permeability) or to restore EC barrier integrity via peripheral cytoskeletal remodeling and formation of cytoskeletal-driven lamellipodia and focal adhesions that promote EC gap closure. The analysis of these biophysical events represent the thematic underpinnings of this PPG and will be conducted by an outstanding translational team of gifted and interactive basic and physician-scientist investigators who will utilize clinically relevant bioactive/biophysical stimuli (VEGF, TNFα, LPS, mechanical stress) both in vitro and in preclinical models of ARDS and VILI. Project #1 will perform sophisticated structure, function and genetic analyses of the multi-functional non-muscle myosin light chain kinase (nmMLCK) isoform in the context of paracellular gap regulation and as a target for lung vascular barrier restoration. As EC barrier-regulatory enhancement is determined by peripheral actin remodeling, Project #2 will provide novel information regarding the interactions between nmMLCK and key actin-binding proteins (cortactin, Ena/VASP-like or EVL, c-Abl) in EC barrierrestorative responses (peripheral cytoskeletal remodeling, lamellipodial dynamics, gap closure). Focal adhesion (FA) proteins are complex participants in both the pathobiology and resolution of ARDS via bidirectional signaling to the cytoskeleton. Project #3 will interrogate the role of the FA proteins, integrin β4 and paxillin, in cytoskeletal linkages to focal adhesion dynamics (assembly/disassembly) and lamellipodialmediated closure of inflammation-induced EC gaps as well as the influence of single nucleotide polymorphisms (SNPs) and post-translational modifications (PTMs) on FA structure /function. Supported by four highly interactive cores, our programmatic approaches are woven into unique PPG features: i) testing of novel ARDS/VILI therapies designed to attenuate the highly druggable lung EC permeability pathway (MLCK inhibitors, S1P, HGF, integrin β4 antibodies) and, ii) the interrogation of ARDS-associated SNPs in key PPG EC barrier-regulatory genes, studies of enormous importance as African descent individuals are a population at high risk for reduced survival in ARDS. By leveraging the outstanding scientific environment at the University of Arizona and long standing collaborations with University of Illinois scientists, this "model translational PPG" is exceptionally suited to provide comprehensive mechanistic understanding of lung vascular barrier regulation, and facilitate the development of therapeutic targets to restore the integrity of the injured pulmonary circulation.

描述（由申请人提供）： 修订后的 A1 PPG 应用仍然是高度转化的 PPG，重点关注内皮细胞 (EC) 细胞骨架在急性呼吸窘迫综合征 (ARDS) 和呼吸机引起的肺损伤 (VILI)（一种破坏性疾病）的病理学中的关键作用。死亡率约为 30-35%，是由肺血管通透性增加决定的，这是对 ARDS 死亡率的主要影响。我们提出的临床相关 PPG 研究将确定其分子机制。 EC信号传导驱动细胞骨架重塑，通过产生细胞旁间隙（增加肺血管通透性）破坏血管完整性，或通过外周细胞骨架重塑和形成细胞骨架驱动的片状伪足和促进EC间隙闭合的粘着斑来恢复EC屏障完整性。对这些生物物理事件的分析代表了本 PPG 的主题基础，并将由一支由才华横溢、互动的基础研究人员和医师科学家组成的杰出翻译团队进行谁将利用临床相关的生物活性/生物物理刺激（VEGF、TNFα、LPS、 项目 #1 将在 ARDS 和 VILI 的体外和临床前模型中对细胞旁间隙调节的多功能非肌肉肌球蛋白轻链激酶 (nmMLCK) 亚型进行复杂的结构、功能和遗传分析。由于 EC 屏障调节增强是由外周肌动蛋白重塑决定的，因此项目 #2 将提供有关 nmMLCK 和关键肌动蛋白结合蛋白之间相互作用的新信息。 EC 屏障恢复反应（外周细胞骨架重塑、板状足动力学、间隙闭合）中的（cortactin、Ena/VASP 样或 EVL、c-Abl）蛋白是 ARDS 病理生物学和通过双向解决的复杂参与者。项目 #3 将探究 FA 蛋白、整合素 β4 和桩蛋白在细胞骨架中的作用。细胞骨架与粘着斑动力学（组装/拆卸）和片状足介导的炎症诱导的 EC 间隙闭合的联系，以及单核苷酸多态性 (SNP) 和翻译后修饰 (PTM) 对 FA 结构/功能的影响 由四个支持。高度互动的核心，我们的程序化方法高度融入独特的 PPG 功能：i) 测试旨在减弱药物作用的新型 ARDS/VILI 疗法肺 EC 通透性通路（MLCK 抑制剂、S1P、HGF、整合素 β4 抗体）以及 ii) 关键 PPG EC 屏障调节基因中与 ARDS 相关的 SNP 的询问，这些研究非常重要，因为非洲人后裔是高危人群通过利用亚利桑那大学出色的科学环境以及与伊利诺伊大学科学家的长期合作，这种“模型转化 PPG”特别适合提供全面的治疗。了解肺血管屏障调节的机制，并促进治疗靶点的开发，以恢复受损肺循环的完整性。