Microtubule involvement in lung endothelial pathobiology

微管参与肺内皮病理学

• 批准号: 7347546
• 负责人: ALEXANDER D VERIN
• 金额: $ 43.97万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7347546
• 关键词: Actins; Actomyosin; Affect; Agonist; Attenuated; Binding Proteins; Calmodulin; Complex; Condition; Cytoskeletal Proteins; Cytoskeleton; DNA Sequence Rearrangement; Data; Disruption; Endothelial Cells; Endothelium; Enzymes; Figs - dietary; Functional disorder; LIM Domain Kinase 1; Link; Lung; Lung Inflammation; MAP Kinase Gene; MAPK14 gene; Mechanical Stimulation; Mediating; Membrane; Microfilaments; Microtubule Alteration; Microtubule Stabilization; Microtubule-Associated Proteins; Microtubules; Modeling; Molecular; Myosin Light Chain Kinase; Myosin Light Chains; Myosin Phosphatase Pathway; Pathway interactions; Permeability; Phosphorylation; Phosphotransferases; Protein Isoforms; Protein Kinase; Proteins; Receptor Signaling; Regulation; Rho-associated kinase; Role; Signal Pathway; Signal Transduction; Stress Fibers; Stretching; Structure; Thrombin; Time; Tubulin; calmodulin-dependent protein kinase II; cofilin; filamin; inhibitor/antagonist; myosin phosphatase; novel; programs; receptor; rho; sphingosine 1-phosphate; tau Proteins

Disturbances in endothelial cell (EC) barrier regulation is a hallmark of lung inflammation and are critically dependent upon rearrangements in the EC cytoskeleton (microfilaments and microtubules). We have previously shown the involvement of actomyosin-driven microfilament contraction and increase in myosin light chain (MLC) phosphorylation in the receptor agonists-mediated permeability. In contrast, information about the role of microtubule network in EC barrier regulation is limited. Our novel observations indicate that thrombin- and TGFb-mediated EC barrier dysfunction is dependent (in part) upon microtubule remodeling as stabilization of microtubule network significantly attenuates thrombin- and TGFb-induced permeability. Our data demonstrate strong correlation between agonist-induced microtubule dissolution and increased phosphorylation of several microtubule-associated proteins (MAPs) including calmodulin (CaM)-dependent kinase II (CaMKII) targets, tau and filamin, implicating a possible role of MAPs and CaM-dependent enzymes such as CaMKII and MLC kinase (MLCK)in receptor agonist-mediated microtubule remodeling. We have recently shown that microtubule inhibitors significantly increases EC MLC phosphorylation and permeability indicating the importance of microtubules in maintaning the EC barrier. Barrier dysfunction induced by microtubule dissolution is significantly attenuated by Rho and p38 MAP kinase pathways inhibition. Together, these data clearly convey the complex involvement of microtubule-mediated pathways in the regulation of EC permeability. The overall objective of this proposal is to examine the molecular mechanisms by which microtubules contribute to the receptor agonist-mediated EC barrier dysfunction under static and cyclic stretch-stimulating conditions. SA#1 will examine the role of CaMKII in thrombin and TGFb-induced microtubule dissolution, SA#2 will characterize the involvement of MLCK in agonist-mediated microtubule remodeling, SA#3 will examine the role of Rho pathway in microtubule-mediated increases in MLC phosphorylation and permeability, SA#4 will identify p38 MAPK pathways involved in microtubulemediatedEC contractility and barrier dysfunction, SA#5 will established the link between sphingosine 1-phosphate barrier-protective signaling and receptor agonist-mediated microtubule dissolution. These studies will provide an understanding of novel signaling pathways involved in lung EC barrier regulation.

内皮细胞 (EC) 屏障调节紊乱是肺部炎症的一个标志，并且严重依赖于 EC 细胞骨架（微丝和微管）的重排。我们之前已经证明，肌动球蛋白驱动的微丝收缩和肌球蛋白轻链（MLC）磷酸化的增加参与了受体激动剂介导的通透性。相比之下，有关微管网络在 EC 屏障调节中的作用的信息有限。我们的新观察表明，凝血酶和 TGFb 介导的 EC 屏障功能障碍（部分）依赖于微管重塑，因为微管网络的稳定显着减弱了凝血酶和 TGFb 诱导的通透性。我们的 数据表明，激动剂诱导的微管溶解与几种微管相关蛋白 (MAP) 的磷酸化增加之间存在很强的相关性，包括钙调蛋白 (CaM) 依赖性激酶 II (CaMKII) 靶标、tau 和细丝蛋白，这表明 MAP 和 CaM 依赖性酶（例如 CaMKII 和 MLC 激酶 (MLCK)）在受体中可能发挥作用 激动剂介导的微管重塑。我们最近表明，微管抑制剂显着增加 EC MLC 磷酸化和通透性，表明微管在维持 EC 屏障方面的重要性。 Rho 和 p38 MAP 激酶途径抑制可显着减弱微管溶解引起的屏障功能障碍。总之，这些数据清楚地传达了微管介导的途径在 EC 通透性调节中的复杂参与。该提案的总体目标是检查分子 微管在静态和循环拉伸刺激条件下导致受体激动剂介导的 EC 屏障功能障碍的机制。 SA#1 将检查 CaMKII 在凝血酶和 TGFb 诱导的微管溶解中的作用，SA#2 将描述 MLCK 在激动剂介导的微管重塑中的作用，SA#3 将检查 Rho 通路在微管介导的 MLC 磷酸化和通透性增加中的作用，SA#4 将识别参与微管介导的微管重塑的 p38 MAPK 通路 微管介导的 EC 收缩性和屏障功能障碍，SA#5 将建立 1-磷酸鞘氨醇屏障保护信号和受体激动剂介导的微管溶解之间的联系。这些研究将有助于了解参与肺 EC 屏障调节的新信号通路。