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: 8220907
• 负责人: Sanda A Predescu
• 金额: $ 36.63万
• 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 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8220907
• 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）：肺小泡和肺内皮细胞（ECs）功能异常可能导致肺血管通透性改变。这可能是急性肺损伤背后复杂病理生理的关键因素。调控小泡介导的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。我们的支持数据表明，肺内皮细胞中至关重要的支架蛋白，交叉蛋白-1s (ITSN-1s)，通过与动力蛋白和其他调节内皮通透性的关键蛋白的相互作用，在小泡运输中发挥重要作用。我们的支持数据表明，itsn -1对于在小窝颈部招募动力蛋白至关重要，动力蛋白在内吞部位的存在及其水解GTP的能力对于小窝胞吞过程中从质膜释放出来至关重要。然而，itsn -1与动力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不明确。引人注目的是，缺乏itsn -1的内皮细胞通过激活线粒体死亡途径而发生凋亡，其机制涉及存活激酶Erk1/2的失活。因此，itsn -1除了调节胞吸外，也是一种抗凋亡蛋白，可能在介导ECs凋亡继发性肺微血管损伤中发挥作用。提出的研究的中心假设是，itsn -1是一种关键蛋白，介导小泡内吞作用与调节肺血管内皮通透性和EC存活的信号通路之间的串扰。以下具体目标将验证这一假设：具体目标1将通过其SH3结构域定义itsn -1作为动力蛋白功能调节剂的作用。特异性目的2将探讨itsn -1 /dynamin相互作用在小鼠肺微血管内皮细胞质膜小泡释放机制和调节肺微血管通透性中的功能意义。特异性Aim 3将描述itsn -1控制的保护肺内皮免受凋亡细胞死亡和肺血管内皮损伤的信号机制。随着目标的成功完成，我们希望对以下方面提供新的见解：i) ITSN-1的SH3结构域在肺微血管内皮细胞中调节动力蛋白功能中的关键作用；ii) ITSN/动力蛋白相互作用在小泡内吞作用和肺内皮屏障功能中的功能意义；iii) ITSN-1在内皮细胞中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后果。我们希望对胞吞作用及其对肺血管内皮通透性的调节提供新的认识，并针对肺微血管屏障的不适当渗漏提供特定的分子治疗靶点。本研究将在以下方面提供新的见解：i)交叉蛋白-1s在调节肺血管内皮通透性中的作用；ii)交叉蛋白-1s在内皮细胞中抗凋亡作用的信号通路和机制。证实了交叉蛋白-1在微血管通透性、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，对抗肺微血管屏障的不适当渗漏，减缓急性呼吸窘迫综合征（ARDS）和急性肺损伤（ALI）相关的肺泡破坏。

项目成果

Endocytic deficiency induced by ITSN-1s knockdown alters the Smad2/3-Erk1/2 signaling balance downstream of Alk5.

• DOI: 10.1242/jcs.163030
• 发表时间: 2015-04-15
• 期刊: Journal of cell science
• 影响因子: 4
• 作者: Bardita C;Predescu DN;Sha F;Patel M;Balaji G;Predescu SA
• 通讯作者: Predescu SA

Pro-inflammatory endothelial cell dysfunction is associated with intersectin-1s down-regulation.

• DOI: 10.1186/1465-9921-12-46
• 发表时间: 2011-04-12
• 期刊: Respiratory research
• 影响因子: 5.8
• 作者: Singla S;Predescu D;Bardita C;Wang M;Zhang J;Balk RA;Predescu S
• 通讯作者: Predescu S

Impaired caveolae function and upregulation of alternative endocytic pathways induced by experimental modulation of intersectin-1s expression in mouse lung endothelium.

• DOI: 10.1155/2012/672705
• 发表时间: 2012
• 期刊: Biochemistry research international
• 影响因子: 3
• 作者: Predescu DN;Neamu R;Bardita C;Wang M;Predescu SA
• 通讯作者: Predescu SA

Rac1-mediated cytoskeleton rearrangements induced by intersectin-1s deficiency promotes lung cancer cell proliferation, migration and metastasis.

• DOI: 10.1186/s12943-016-0543-1
• 发表时间: 2016-09-14
• 期刊: Molecular cancer
• 影响因子: 37.3
• 作者: Jeganathan N;Predescu D;Zhang J;Sha F;Bardita C;Patel M;Wood S;Borgia JA;Balk RA;Predescu S
• 通讯作者: Predescu S

In vivo knockdown of intersectin-1s alters endothelial cell phenotype and causes microvascular remodeling in the mouse lungs.

• DOI: 10.1007/s10495-012-0762-x
• 发表时间: 2013-01
• 期刊: APOPTOSIS
• 影响因子: 7.2
• 作者: Bardita, Cristina;Predescu, Dan;Justice, Matthew J.;Petrache, Irina;Predescu, Sanda
• 通讯作者: Predescu, Sanda