Mechanobiology of Lung Fibrosis

肺纤维化的力学生物学

• 批准号: 9906248
• 负责人: Daniel J. Tschumperlin
• 金额: $ 56.95万
• 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 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9906248
• 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; Epithelium; 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; 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.

项目摘要 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传导促进疾病 进展。我们发布的初步数据表明，Yap和Taz，转录效应 河马途径是IPF中成纤维细胞激活的关键调节剂，并控制ECM沉积和 通过成纤维细胞僵硬。但是，YAP和TAZ是多个途径的下游，并且在 多种肺部细胞类型，使针对它们理论的努力变得复杂。因此，我们将重点放在这里 开发一种以成纤维细胞为目标的方法来抑制YAP/TAZ。具体来说，我们已经确定了GPCR 通过GαS耦合多巴胺D1受体（DRD1）作为成纤维细胞选择性方法的激动剂 失活的Yap和Taz。我们的体外和体内初步数据证明了药物刺激 DRD1不仅会减弱成纤维细胞的激活，而且在功能上将其状态从矩阵沉积到 基质降解和矩阵加强的逆转。这些响应取决于YAP/TAZ的抑制作用 它们在表达组成性活性TAZ突变蛋白的细胞中丢失。已发表的报告表明 内源性多巴胺能信号传导存在于正常肺中。我们的初步数据表明 在实验纤维化过程中，多巴胺合成途径在小鼠实验性纤维化过程中瞬时降低，并稳定降低 在患有IPF的受试者的肺中。这是我们海报的中心假设，即多巴胺信号通常 促进肺损伤后的纤维化分辨率，在IPF中丢失，并且可以由DRD1激动剂瞄准 逆转实验性肺纤维化。我们建议以三个特定目的检验这一假设，结合 肺上皮细胞和多巴胺能信号对肺的多巴胺合成的体外分析 成纤维细胞功能，以及内源性多巴胺的实验纤维化的体内分析 生产丢失或外源性增强的药物。拟议的研究将共同​​描述 一种新型的受体介导的机制，可以将成纤维细胞从促进纤维化切换为纤维化 解决状态，测试该途径在耐用纤维化模型中的外源靶向的治疗效率， 并探索该途径的内源性活动​​是否通常可以保护并解决渐进性 纤维化，在人类疾病中丧失。