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; 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）： 该修订后的A1 PPG应用仍然是高度翻译的PPG，重点是内皮细胞（EC）细胞骨架在急性呼吸遇险综合征（ARDS）和呼吸机诱导的肺损伤（VILI）中的关键作用。 ARDS是一种毁灭性疾病，死亡率约为30-35％，是由肺血管通透性的增加来定义的，这对ARDS死亡率产生了重大影响。我们提出的与临床相关的PPG研究将定义EC信号传导通过产生副细胞间隙（增加肺血管通透性）或通过外外外围细胞骨架的EC Barrier恢复EC Barrier的整体重塑，使细胞骨架的分子机制驱动细胞骨架重塑，以破坏血管完整性（增加肺血管通透性）或恢复。以及促进EC缝隙封闭的局灶性粘连。对这些生物物理事件的分析代表了该PPG的主题基础，并将由一支出色的天赋和互动基础和物理科学家研究人员进行，他们将利用临床上相关的生物活性/生物物理刺激（VEGF，VEGF，TNFα，TNFα，TNFα，LPS，LPS，LPS，LPS，LPS，LPS，LPS，LPS，LPS，LPS，LPS，LPS，LPS，LPS 机械应力）在体外和ARD和VILI的临床前模型中均应。项目＃1将在细胞细胞间隙调节的背景下进行多功能非肌肉肌球蛋白轻链激酶（NMMLCK）同工型进行复杂的结构，功能和遗传分析，并作为肺血管屏障恢复的靶标。 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).局灶性粘附（FA）蛋白是通过双向信号传导向细胞骨架的双向信号传导的病理生物学和分辨率的复杂参与者。项目＃3将询问FA蛋白，整联蛋白β4和帕克西蛋白的作用，在局部骨架上与局部粘附动力学（组装 /拆卸）以及发炎诱导的EC差距的封闭以及单核肽多态性（Snps-snps-snps-snps-snpers-ptrans-pats-pta insivic intivic intivic of）的影响（组装 /拆卸）和lamellipodialipialipialdialipialdialipialdialdial诱导的闭合。 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 drugable 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基因，随着非洲血统个体的人群的高度降低ARDS的人口，其重要性的研究提高了。通过利用亚利桑那大学的杰出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”非常适合对肺血管屏障调节的全面机械理解，并促进了治疗靶标的发展，以恢复受伤肺循环的良好的完整性。