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）屏障调节中的干扰是肺部炎症的标志，并且严重取决于EC细胞骨架（微丝和微管）的重排。以前，我们已经显示了肌动蛋白驱动的微丝收缩和肌球蛋白轻链（MLC）磷酸化的涉及受体激动剂介导的渗透性。相反，有关微管网络在EC屏障调节中的作用的信息有限。我们的新观察结果表明，凝血酶和TGFB介导的EC屏障功能障碍（一定程度地）取决于微管重塑，因为微管网络的稳定性显着减弱了凝血酶和TGFB诱导的渗透性。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）的磷酸化之间的牢固相关性，包括钙调蛋白（CAM） - 依赖性激酶II（CAMKII）靶标，TAU和丝胺，与camkii和MLCKKII和MLCKKII的可能作用有关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磷酸化和渗透性，表明微管在维持EC屏障中的重要性。由微管溶解引起的屏障功能障碍被RHO和p38 MAP激酶途径抑制显着减弱。总之，这些数据清楚地表达了微管介导的途径在EC渗透性调节中的复杂涉及。该提案的总体目的是检查分子 在静态和环状拉伸刺激条件下，微管有助于受体激动剂介导的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将表征MLCK参与激动剂介导的微管重塑中的SA＃3，SA＃3将检查Rho Pathway在Milotubule介导的Milotubule介导的MLC磷酸化和SA＃4的作用，并识别SA＃4微管介导的收缩性和屏障功能障碍，SA＃5将建立鞘氨醇1-磷酸屏障保护信号传导与受体激动剂介导的微管溶解之间的联系。这些研究将提供对肺部屏障调节中涉及的新型信号通路的理解。