Differential mechano-signaling in vascular endothelium by varying degrees of mechanical stretch - Resubmission 01

通过不同程度的机械拉伸在血管内皮中产生差异性机械信号 - 重新提交 01

基本信息

批准号：
9280991
负责人：
Anna Birukova
金额：
$ 38.63万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2020-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9280991
关键词：
Adaptor Signaling Protein Address Adherens Junction Blood Vessels Cells Cellular Mechanotransduction Characteristics Complex Data Down-Regulation Drug Targeting Edema Endothelial Cells Endothelium Environment Environmental air flow Event Exposure to Extravasation Functional disorder Guanine Nucleotide Exchange Factors Guanosine Triphosphate Phosphohydrolases Heart failure Hyperemia Hypoxia Inflammation Inflammatory Intercellular Junctions Intervention Lead Leukocytes Life Liquid substance Lung Mechanical Stress Mechanical ventilation Mechanics Microcirculatory Bed Microvascular Permeability Molecular Monomeric GTP-Binding Proteins Names Nucleotides Nutrient Organ Pathologic Pathology Pathway interactions Periodicity Permeability Physiological Pilot Projects Play Property Pulmonary Edema Pulmonary Hypertension Recovery Recruitment Activity Redness Regulation Role Signal Pathway Signal Transduction Specific qualifier value Stretching Stroke Swelling Testing Tidal Volume Tight Junctions Time Tissues Variant Vascular Endothelial Cell Vascular Endothelium Vascular Permeabilities Ventilator-induced lung injury in vivo mechanical force mechanotransduction mouse model novel preconditioning response restoration rho rho GTP-Binding Proteins tumor

项目摘要

Differential mechano-signaling in vascular endothelium by varying degrees of mechanical stretch Excessive mechanical forces imposed on the microvascular endothelium lead to increased microvascular permeability accompanying life-threatening conditions such as stroke, pulmonary hypertension, or ventilator induced lung injury, to name a few. We have previously characterized signaling pathways induced in vascular endothelium by high magnitude cyclic stretch (CS) and described a key role of Rho GTPase in high CS- induced endothelial barrier dysfunction. However, cellular mechanotransduction complexes which transform mechanical signals to cellular responses remain to be characterized. Our unpublished pilot studies indicate that Rap1 signaling is activated by physiologically relevant low magnitude (5%) CS and promotes re-assembly of tight junctions disrupted by cell preconditioning at pathologically relevant high magnitude (18%) CS. The preliminary data also suggest that low CS-induced EC barrier restoration is associated with accumulation of adaptor protein cingulin at the tight junctions and formation of cingulin-GEF-H1 complex. The central hypothesis tested in this application is that recovery of vascular endothelial barrier after pathologic mechanical stress may be accelerated by cell exposure to physiologic CS levels and involves Rap1-dependent reassembly of endothelial tight junctions, recruitment of cingulin to the tight junctions, and stimulation of cingulin - GEF-H1 interaction. These events lead to inhibition of GEF-H1 nucleotide exchange activity, suppression of Rho- dependent barrier disruptive mechanisms, and accelerated recovery of the vascular endothelial barrier. The following questions will be addressed: Aim-1 will study cingulin-dependent mechanisms of endothelial barrier regulation by physiologic mechanical stretch. Aim-2 will study the role of cingulin in downregulation of the Rho pathway in mechanically stimulated microvascular endothelium after a switch from pathologic to physiologic CS amplitude. Aim-3 will evaluate cingulin-dependent vascular protective mechanisms in a mouse model of mechanical ventilation.

通过不同程度的机械拉伸在血管内皮中产生差异性机械信号对微血管内皮施加过大的机械力导致微血管增加伴随危及生命的疾病（如中风、肺动脉高压或呼吸机）的渗透性诱发肺损伤等。我们之前已经表征了血管中诱导的信号通路内皮细胞通过高强度循环拉伸（CS）并描述了 Rho GTPase 在高 CS 中的关键作用诱发内皮屏障功能障碍。然而，细胞力转导复合物可转化细胞反应的机械信号仍有待表征。我们未发表的试点研究表明 Rap1 信号传导由生理相关的低强度 (5%) CS 激活并促进重新组装细胞预处理在病理相关的高强度 (18%) CS 下破坏紧密连接。这初步数据还表明，低 CS 诱导的 EC 屏障恢复与接头蛋白 cingulin 在紧密连接处并形成 cingulin-GEF-H1 复合物。中央本申请检验的假设是病理机械损伤后血管内皮屏障的恢复细胞暴露于生理性 CS 水平可能会加速应激，并涉及 Rap1 依赖性重组内皮紧密连接的形成、将扣带蛋白募集到紧密连接以及刺激扣带蛋白 - GEF-H1 相互作用。这些事件导致 GEF-H1 核苷酸交换活性受到抑制，Rho- 依赖性屏障破坏机制，并加速血管内皮屏障的恢复。这将解决以下问题： Aim-1 将研究内皮屏障的 cingulin 依赖性机制通过生理机械拉伸进行调节。 Aim-2 将研究 cingulin 在 Rho 下调中的作用从病理性转变为生理性后机械刺激微血管内皮的通路 CS 幅度。 Aim-3 将在小鼠模型中评估 cingulin 依赖性血管保护机制机械通气。