Intersectin-1s regulates lung vascular permeability and endothelial cell survival

Intersectin-1s 调节肺血管通透性和内皮细胞存活

• 批准号: 7579028
• 负责人: Sanda A Predescu
• 金额: $ 37万
• 依托单位: RUSH UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7579028
• 关键词: Acute; Acute Lung Injury; Address; Adult Respiratory Distress Syndrome; Albumins; Apoptosis; Apoptotic; Binding Sites; Biochemical; Blood Vessels; Caveolae; Cell Death; Cell Survival; Cell membrane; Cessation of life; Characteristics; Chemicals; Coiled-Coil Domain; Complement; Complex; Data; Down-Regulation; Dynamin; Electron Microscopy; Elements; Endocytosis; Endothelial Cells; Endothelium; Figs - dietary; Functional disorder; Guanosine Triphosphate; Guanosine Triphosphate Phosphohydrolases; Hydrolysis; Impairment; Injury; Link; Lung; Lung diseases; Mediating; Microvascular Permeability; Mitochondria; Molecular; Molecular Biology; Mus; Neck; Pathway interactions; Permeability; Phosphorylation; Phosphotransferases; Play; Proline-Rich Domain; Property; Protein Family; Protein Overexpression; Proteins; RNA Interference; Ras/Raf; Recruitment Activity; Regulation; Role; SH3 Domains; Scaffolding Protein; Secondary to; Signal Pathway; Signal Transduction; Site; Structure; Testing; Vascular Permeabilities; Work; alveolar destruction; base; insight; intersectin 1; knock-down; novel; platelet protein P47; protein expression; therapeutic target; trafficking; transcytosis

DESCRIPTION (provided by applicant): Abnormalities in caveolae and lung endothelial cell (ECs) function may result in alterations of lung vascular permeability. This may be a key element of the complex pathophysiology underlying acute lung injury. The understanding of the molecular mechanisms that regulate caveolae-mediated transcytosis and its role in mediating increased lung microvessel endothelial permeability are unclear. Our Supporting Data suggest that the crucial scaffold protein in lung ECs, intersectin-1s (ITSN-1s), plays an important role in caveolae trafficking through its interaction with dynamin and other key proteins regulating endothelial permeability. Our Supporting Data suggest that ITSN-1s is crucial in recruiting dynamin at the neck region of caveolae and that dynamin presence at the endocytic site and its ability to hydrolyze GTP are essential for caveolae release from the plasma membrane during caveolae transcytosis. However, the relationship between ITSN-1s and dynamin and its significance in the mechanism of increased lung vascular endothelial permeability are not yet defined. Strikingly, ECs deficient in ITSN-1s became apoptotic by activation of the mitochondrial death pathway through a mechanism that involves inactivation of the survival kinase Erk1/2. Thus, besides regulating transcytosis, ITSN-1s is also an anti-apoptotic protein that may have consequences in mediating lung microvascular injury secondary to apoptosis of ECs. The central hypothesis of the proposed studies is that ITSN-1s is a pivotal protein mediating the cross-talk between caveolae endocytosis and signaling pathways that regulates lung vascular endothelial permeability and EC survival. The following specific aims will test this hypothesis: Specific Aim 1 will define the role of ITSN-1s, via its SH3 domains, as a regulator of dynamin function. Specific Aim 2 will address the functional significance of ITSN-1s/dynamin interaction in the mechanism of caveolae release from the plasma membrane of lung microvessel ECs and in regulating lung microvascular permeability in mice. Specific Aim 3 will delineate the signaling mechanisms controlled by ITSN-1s that defend lung endothelium against apoptotic cell death and lung vascular endothelial injury. With the successful completion of the aims, we hope to provide novel insights into the i) pivotal role of the SH3 domains of ITSN-1s in regulation of dynamin function in lung microvessel ECs, ii) functional significance of ITSN/dynamin interaction in caveolae endocytosis and lung endothelial barrier function, and iii) signaling pathways and mechanisms governing the anti-apoptotic effects of ITSN-1s in ECs and its pathophysiological consequence in the mechanism of lung microvascular injury. We hope to provide a novel understanding of transcytosis and its regulation of lung vascular endothelial permeability and specific molecular therapeutic targets directed against inappropriate leakiness of the lung microvascular barrier. This study will provide novel insights regarding i) the role of intersectin-1s in regulation of lung vascular endothelial permeability and ii) the signaling pathways and mechanisms governing the anti-apoptotic effects of intersectin-1s in endothelial cells. Confirming intersectin-1s as a link between microvascular permeability, apoptosis and lung disease, we may find new targets against inappropriate leakiness of the lung microvascular barrier and slow the alveolar destruction associated with acute respiratory distress syndrome (ARDS) and acute lung injury (ALI).

描述（由申请方提供）：小窝和肺内皮细胞（EC）功能的缺失可能导致肺血管通透性的改变。这可能是急性肺损伤复杂病理生理学的关键因素。调节小窝介导的转胞吞作用的分子机制及其在介导肺微血管内皮通透性增加中的作用尚不清楚。我们的支持性数据表明，肺EC中的关键支架蛋白，interstin-1 s（ITSN-1 s），通过与发动蛋白和其他调节内皮通透性的关键蛋白的相互作用，在小窝运输中发挥重要作用。我们的支持性数据表明，ITSN-1 s是至关重要的招募发动蛋白在颈部区域的小窝和发动蛋白的存在，在内吞位点和其水解GTP的能力是必不可少的小窝释放从质膜在小窝转胞吞。然而，ITSN-1 s和发动蛋白之间的关系及其在肺血管内皮通透性增加机制中的意义尚未明确。引人注目的是，缺乏ITSN-1 s的内皮细胞通过激活线粒体死亡途径而凋亡，该途径通过一种涉及生存激酶Erk 1/2失活的机制。因此，除了调节转胞吞作用，ITSN-1 s也是一种抗凋亡蛋白，其可能在介导继发于EC凋亡的肺微血管损伤中具有后果。所提出的研究的中心假设是ITSN-1 s是介导小窝内吞作用和调节肺血管内皮通透性和EC存活的信号通路之间的串扰的关键蛋白。以下具体目标将测试这一假设：具体目标1将定义ITSN-1的作用，通过其SH 3结构域，作为动力蛋白功能的调节剂。具体目标2将阐述ITSN-1 s/发动蛋白相互作用在小鼠肺微血管内皮细胞质膜小窝释放机制和调节肺微血管通透性中的功能意义。具体目标3将描述由ITSN-1控制的信号传导机制，其保护肺内皮免受凋亡性细胞死亡和肺血管内皮损伤。随着目标的成功完成，我们希望提供新的见解i）ITSN-1的SH 3结构域在调节肺微血管EC中动力蛋白功能中的关键作用，ii）ITSN/动力蛋白相互作用在小窝内吞和肺内皮屏障功能中的功能意义，以及iii）控制ITSN-1在EC中的抗凋亡作用的信号通路和机制及其在肺微血管损伤机制中的病理生理学后果。我们希望提供一个新的理解转胞吞及其调节肺血管内皮细胞通透性和特定的分子治疗靶点针对不适当的泄漏肺微血管屏障。本研究将提供新的见解i）intersectin-1 s在调节肺血管内皮通透性的作用和ii）intersectin-1 s在内皮细胞中的抗凋亡作用的信号通路和机制。将intersectin-1 s作为微血管通透性、细胞凋亡和肺部疾病之间的联系，我们可能会发现新的靶点，以防止肺微血管屏障的不适当泄漏，并减缓与急性呼吸窘迫综合征（ARDS）和急性肺损伤（ALI）相关的肺泡破坏。