Mechanobiology of Lung Fibrosis

肺纤维化的力学生物学

• 批准号: 10390336
• 负责人: Daniel J. Tschumperlin
• 金额: $ 56.66万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-06 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10390336
• 关键词: Acute; Agonist; Area; Attenuated; Biochemical Feedback; Biomechanics; Bleomycin; Cell physiology; Cells; Coculture Techniques; Collagen; Coupled; DOPA decarboxylase; DRD1 gene; Data; Degradation Pathway; Deposition; Depressed mood; Disease; Disease Progression; Dopamine; Dopamine D1 Receptor; Dopamine Receptor; Enzymes; Epithelial; Epithelial Cells; Evaluation; Extracellular Matrix; Fibroblasts; Fibrosis; G-Protein-Coupled Receptors; GTP-Binding Protein alpha Subunits, Gs; Genetic; Genetic Transcription; Homeostasis; In Vitro; Injury; LATS1 gene; Lung; MMP14 gene; Mediating; Mesenchymal; Modeling; Molecular; Mus; Nature; Organoids; Pathologic; Pathway interactions; Peptide Hydrolases; Pharmacology; Play; Production; Proteins; Publishing; Pulmonary Fibrosis; Regulation; Reporting; Resolution; Role; Signal Pathway; Signal Transduction; Testing; Therapeutic; Treatment Efficacy; aged; antagonist; cell type; cellular targeting; crosslink; efficacy testing; experimental analysis; experimental study; human disease; in vivo; in vivo evaluation; injury and repair; lung injury; mutant; novel; novel strategies; programs; receptor; regenerative; response; therapeutic evaluation; therapeutic target

Project Summary Pulmonary fibrosis is a progressive and ultimately fatal disease in which ongoing extracellular matrix (ECM) deposition and feedback biochemical and biomechanical signaling from this matrix promotes disease progression. Our published and preliminary data demonstrate that YAP and TAZ, transcriptional effectors of the Hippo pathway, are pivotal regulators of fibroblast activation in IPF, and control both ECM deposition and stiffening by fibroblasts. However, YAP and TAZ are downstream of multiple pathways, and play critical roles in multiple lung cell types, complicating efforts to target them therapeutically. Therefore we focus here on developing a fibroblast-targeted approach to YAP/TAZ inhibition. Specifically, we have identified GPCR agonism via Gαs-coupled dopamine D1 Receptor (DRD1) as a fibroblast selective approach through which to inactivate YAP and TAZ. Our in vitro and in vivo preliminary data demonstrate that pharmacologic stimulation of DRD1 not only attenuates fibroblast activation, but functionally reverses their state from matrix depositing to matrix degradation and reversal of matrix stiffening. These responses depend on inhibition of YAP/TAZ, as they are lost in cells expressing constitutively active TAZ mutant protein. Published reports suggest that endogenous dopaminergic signaling is present in the normal lung; our preliminary data demonstrate that the dopamine synthetic pathway is transiently depressed during experimental fibrosis in mice, and stably reduced in the lungs of subjects with IPF. Thus, we posit the central hypothesis that dopamine signaling normally promotes fibrosis resolution after lung injury, is lost in IPF, and can be selectively targeted by DRD1 agonism to reverse experimental lung fibrosis. We propose to test this hypothesis in three specific aims, combining in vitro analysis of dopamine synthesis by lung epithelial cells and dopaminergic signaling effects on lung fibroblast function, as well as in vivo analysis of experimental fibrosis in mice in which endogenous dopamine production is lost, or exogenously augmented pharmacologically. Together the proposed studies will delineate a novel receptor mediated mechanism by which fibroblast can be switched from fibrosis promoting to fibrosis resolving states, test the therapeutic efficacy of exogenous targeting of this pathway in durable fibrosis models, and explore whether the endogenous activity of this pathway normally protects from and resolves progressive fibrosis, and is lost in human disease.

项目概要 肺纤维化是一种进行性且最终致命的疾病，其中持续的细胞外基质（ECM） 来自该基质的沉积和反馈生化和生物力学信号促进疾病 进展。我们发表的初步数据表明，YAP 和 TAZ，转录效应子 Hippo 通路是 IPF 中成纤维细胞活化的关键调节因子，并控制 ECM 沉积和 成纤维细胞硬化。然而，YAP 和 TAZ 是多种途径的下游，并在 多种肺细胞类型，使针对它们的治疗工作变得复杂。因此我们在这里关注 开发一种针对 YAP/TAZ 抑制的成纤维细胞靶向方法。具体来说，我们已经确定了 GPCR 通过 Gαs 偶联多巴胺 D1 受体 (DRD1) 的激动作用作为成纤维细胞选择性方法，通过该方法 灭活 YAP 和 TAZ。我们的体外和体内初步数据表明，药理刺激 DRD1 的表达不仅减弱成纤维细胞的活化，而且在功能上将其状态从基质沉积逆转为 基体退化和基体硬化逆转。这些反应取决于 YAP/TAZ 的抑制，如 它们在表达组成型活性 TAZ 突变蛋白的细胞中丢失。已发表的报告表明 正常肺中存在内源性多巴胺能信号传导；我们的初步数据表明 小鼠实验性纤维化过程中多巴胺合成途径短暂抑制，并稳定减少 存在于 IPF 受试者的肺部。因此，我们提出的中心假设是多巴胺信号传导正常 促进肺损伤后纤维化消退，在 IPF 中丢失，并且可以通过 DRD1 激动选择性地靶向 逆转实验性肺纤维化。我们建议在三个具体目标中检验这一假设，结合 肺上皮细胞合成多巴胺及多巴胺能信号对肺的影响的体外分析 成纤维细胞功能，以及内源性多巴胺小鼠实验性纤维化的体内分析 生产损失，或药理上外源性增加。拟议的研究将共同​​描述 一种新的受体介导机制，使成纤维细胞从促进纤维化转变为纤维化 解决状态，测试在持久纤维化模型中外源性靶向该途径的治疗效果， 并探讨该途径的内源性活性是否通常能够预防和解决进行性进展 纤维化，并在人类疾病中消失。