Mechanotransduction analysis in a microengineered lung-on-a-chip

微工程肺芯片中的力传导分析

• 批准号: 8862797
• 负责人: DONALD E INGBER
• 金额: $ 61.94万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8862797
• 关键词: Adhesions; Alveolar; Animals; Binding; Biochemistry; Blood Vessels; Blood capillaries; Breathing; Cells; Cellular Mechanotransduction; Cellular biology; Chemicals; Chemotherapy-Oncologic Procedure; Clinical; Coagulation Process; Cytoskeleton; Development; Devices; Disease; Dose-Limiting; Endothelial Cells; Engineering; Environmental air flow; Etiology; Event; Extracellular Matrix; Extravasation; FDA approved; Fibrin; Functional disorder; Goals; Growth and Development function; Health; Heart failure; Human; In Vitro; Inflammation; Inflammatory; Integral Membrane Protein; Integrins; Interleukin-2; Ion Channel; Knowledge; Laboratories; Lead; Life; Lung; Lung diseases; Mechanics; Mediating; Membrane; Microfluidic Microchips; Modeling; Molecular; Molecular Probes; Molecular Target; Motion; Mus; Natural regeneration; Organ; Peptides; Perfusion; Pharmaceutical Preparations; Physiological; Prevention; Process; Pulmonary Edema; Pulmonology; Research; Secondary to; Signal Transduction; Site; Stress; Structure; Supporting Cell; Therapeutic; Tissues; Toxic effect; Vanilloid; Vascular Permeabilities; Work; base; capillary; cytokine; design; experience; improved; inhibitor/antagonist; microsystems; millisecond; novel therapeutics; oxygen transport; pressure; prevent; receptor; response; therapeutic target; therapy development; transmission process; validation studies

 DESCRIPTION: The overall goal of this application is to demonstrate the feasibility of using a microengineered `Lung-on-a-Chip" microfluidic device to probe the molecular mechanism of mechano-chemical signaling in the human lung, and to use this knowledge to develop new and improved inhibitors of pulmonary edema development. One of the most rapid (< 5 msec) mechanical signaling events triggered by force transmission from the microenvironment to the cell via their extracellular matrix adhesions involves integrin-dependent activation of the stress-activated membrane ion channel TRPV4, which appears to be critical for the development of many disease processes, including pulmonary edema. The molecular mechanism by which forces applied to integrin mediate this `early- immediate' mechanical signaling response that activates TRPV4 and lead to pulmonary disease is not well understood. To study this process in vitro, we will use a recently developed human Lung-on-a-Chip microfluidic device that contains an artificial alveolar-capillary interface lined by living human lung alveolar and capillary cells hat experiences physiological breathing motions and regenerates a functional vascular permeability barrier in vitro. Importantly, we previously used this microengineered lung chip to show that a specific chemical inhibitor of TRPV4 activity can prevent pulmonary vascular leakage induced by both interleukin-2 and mechanical deformation (breathing motions). In addition, our preliminary results have revealed that the transmembrane protein CD98 binds to both ß1-integrin and TRPV4, and that it is required for mechanical, but not chemical, activation of TRPV4. Thus, in this project, we propose to use our microengineered human Lung- on-a-Chip device to delineate the molecular mechanism by which forces applied to integrins activate TRPV4, and to develop new therapeutics for pulmonary edema that targets this molecular mechanism. The specific aims include: 1) to define the molecular mechanism by which CD98 mediates ß1-integrin-dependent mechanical activation of TRPV4 in human microvascular endothelial cells, 2) to develop peptide modulators of mechanical signaling through TRPV4 that prevent vascular leakage in the lung-on-a-chip pulmonary edema model, and 3) to validate the peptide inhibitors by demonstrating their ability to prevent vascular leakage in an ex vivo mouse pulmonary edema model.

 描述：本应用的总体目标是证明使用微工程“芯片肺”微流体装置探测人肺中机械化学信号传导的分子机制的可行性，并利用这些知识开发新的和改进的肺水肿发展抑制剂。由来自肺的力传输触发的最快速（< 5 毫秒）的机械信号传导事件之一 细胞通过细胞外基质粘附的微环境涉及应激激活膜离子通道 TRPV4 的整合素依赖性激活，这似乎对于许多疾病过程（包括肺水肿）的发展至关重要。施加于整合素的力介导这种激活TRPV4并导致肺部疾病的“早期立即”机械信号反应的分子机制尚不清楚。 为了在体外研究这一过程，我们将使用最近开发的人体芯片肺微流体装置，该装置包含人工肺泡毛细血管界面，内衬活体人类肺泡和毛细血管细胞，这些细胞经历生理呼吸运动并在体外再生功能性血管通透性屏障。重要的是，我们之前使用这种微工程肺芯片来证明特定的化学抑制剂 TRPV4 活性的降低可以防止白细胞介素 2 和机械变形（呼吸运动）引起的肺血管渗漏。此外，我们的初步结果表明，跨膜蛋白 CD98 与 ß1-整合素和 TRPV4 结合，并且它是 TRPV4 的机械激活（而非化学激活）所必需的。因此，在这个项目中，我们建议使用我们的微工程 人类芯片肺装置描绘了施加于整合素的力激活 TRPV4 的分子机制，并开发针对该分子机制的肺水肿新疗法。具体目标包括：1）确定CD98介导人微血管内皮细胞中TRPV4的β1整合素依赖性机械激活的分子机制， 2) 通过 TRPV4 开发机械信号肽调节剂，防止片上肺肺水肿模型中的血管渗漏，以及 3) 通过证明肽抑制剂在离体小鼠肺水肿模型中防止血管渗漏的能力来验证肽抑制剂。