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: 8033762
• 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/8033762
• 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; Cell physiology; Cessation of life; Characteristics; Chemicals; Coiled-Coil Domain; Complement; Complex; Data; Down-Regulation; Dynamin; Electron Microscopy; Elements; Endocytosis; Endocytosis Pathway; Endothelial Cells; Endothelium; Functional disorder; Guanosine Triphosphate; Guanosine Triphosphate Phosphohydrolases; Impairment; Injury; Link; Lung; Lung diseases; MAP Kinase Gene; 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（由申请人提供）：小窝和肺内皮细胞（ECS）功能的异常可能导致肺血管渗透性的改变。这可能是复杂的急性肺损伤基础病理生理学的关键要素。尚不清楚对调节小窝介导的转胞细胞增多的分子机制的理解及其在介导增加的肺微血管内皮渗透性中的作用尚不清楚。我们的支持数据表明，肺ECS中的至关重要的支架蛋白质相交1S（ITSN-1S）通过与Dynamin和其他调节内皮通透性的关键蛋白的相互作用而在口腔运输中起着重要作用。我们的支持数据表明，ITSN-1S对于在小窝的颈部募集动力蛋白以及在内吞位点的螺旋蛋白的存在以及其水解GTP的能力对于在小窝跨囊肿期间从质膜释放而至关重要的能力至关重要。然而，尚未定义ITSN-1S与元素之间的关系及其在肺血管内皮渗透性增加的机理中的重要性。令人惊讶的是，通过涉及生存激酶ERK1/2失活的机制，通过激活线粒体死亡途径的EC被激活通过线粒体死亡途径凋亡。因此，除了调节转胞细胞增多外，ITSN-1S也是一种抗凋亡蛋白，在介导EC凋亡继发的肺微血管损伤方面可能会产生后果。拟议研究的中心假设是ITSN-1S是一种关键蛋白，介导了可爱的内吞作用和信号传导途径之间的串扰，可调节肺血管内皮通透性和EC生存。以下特定目的将检验以下假设：特定目标1将通过其SH3域来定义ITSN-1的作用，作为Dynamin功能的调节剂。具体的目标2将解决ITSN-1S/动力蛋白相互作用在从肺微血管ECS的质膜中释放机理以及调节小鼠肺微血管通透性方面的功能意义。具体目标3将描述由ITSN-1控制的信号传导机制，这些信号机制捍卫肺部内皮免受凋亡细胞死亡和肺血管内皮损伤。随着目标的成功完成，我们希望对ITSN-1的SH3域在肺微血管ECS中的元素功能调节中的I）i）的关键作用提供新的见解在ECS及其在肺微血管损伤机理中的病理生理后果中。我们希望提供对转胞胞菌病的新了解及其对肺血管内皮渗透性的调节和针对肺微血管屏障不当泄漏的特定分子治疗靶标。这项研究将提供有关i）相交1S在调节肺血管内皮渗透性的作用的新见解，以及II）介绍内皮细胞中索塞蛋白-1S的抗凋亡效应的信号传导途径和机制。确认Intersectin-1s是微血管通透性，凋亡和肺部疾病之间的联系，我们可能会发现新的靶标，以防止肺微血管屏障的不当渗漏，并减慢与急性呼吸道遇险综合征（ARDS）和急性肺损伤相关的肺泡破坏。