Epithelial-Mesenchymal Interactions in Fibrosis Resolution

纤维化消退中的上皮-间质相互作用

• 批准号: 10655172
• 负责人: Daniel J. Tschumperlin
• 金额: $ 59.29万
• 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 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10655172
• 关键词: Ablation; Acceleration; Age; Aging; Biochemical; Biological Availability; Biomechanics; Bleomycin; Cells; Chronic; Collagen; DOPA decarboxylase; Data; Deposition; Depressed mood; Disease; Dissociation; Dopamine; Dopamine Agonists; Dopamine D1 Receptor; Dopamine Receptor; Enzymes; Epithelial Cells; Epithelium; Exhibits; Experimental Models; Extracellular Matrix; Feedback; Fibroblasts; Fibrosis; Funding; Genetic Models; Homeostasis; Human; Impairment; Individual; Injury; Ligands; Lung; Mediator; Medical; Mesenchymal; Modeling; Molecular; Mus; Pathologic; Production; Profibrotic signal; Publishing; Pulmonary Fibrosis; Regulation; Resolution; Role; Signal Transduction; Slice; Source; Structure of parenchyma of lung; System; Testing; Therapeutic; Transcript; Work; aged; cell type; epithelial injury; epithelial repair; fibrotic lung; inhibitor; lung injury; lung repair; mouse model; novel therapeutic intervention; pharmacologic; repaired; response; restoration; single cell analysis; single-cell RNA sequencing; small molecule; small molecule inhibitor; tissue repair

Project Summary Pulmonary fibrosis (PF) remains a major and growing medical burden with unsatisfactory therapeutic options that fail to reverse established disease. Our prior work has identified that YAP/TAZ activation in fibroblasts is a central feature of the pathological feedback loop that propagates progression of PF. In the prior funding cycle, we identified Dopamine D1 receptor (D1R) agonism as a strategy to inactivate YAP/TAZ selectively in lung fibroblasts, leading to accelerated resolution of experimental pulmonary fibrosis in mice in part by switching lung fibroblasts from matrix depositing to matrix degrading state. Moreover, we demonstrated that the lungs of individuals with PF exhibit a deficit in expression of DOPA decarboxylase (DDC), the enzyme that catalyzes conversion of L-DOPA into bioactive dopamine. These studies lead us to propose that restoration of endogenous local dopamine levels in the lung is an essential trigger for the matrix degradation and fibrosis clearance that is essential to successful repair of the lung, but is impaired in PF. Our preliminary data show that Ddc transcripts are transiently depressed in lung tissue of young mice following bleomycin injury and rise during fibrosis resolution, whereas aged mice exhibited sustained reductions in Ddc that parallel persistent fibrosis. Moreover, small molecule inhibition of Ddc enzymatic activity or the D1R from day 21 to 42 post- bleomycin in young mice ablates the spontaneous resolution of lung fibrosis, demonstrating the essential role for dopamine signaling in fibrosis resolution. In addition, we find that dopamine is detectable in supernatants of precision cut lung slices and is diminished in slices cultured from fibrotic lungs, confirming the local synthesis of dopamine within the lung. Based on these findings, we propose to test the central hypothesis that epithelial dopamine synthesis is essential to fibrosis resolution and that restoration of normal dopamine levels in PF lung tissue can promote collagen resorption and repair of the lung. We will test this hypothesis in three aims spanning non-resolving mouse models of pulmonary fibrosis as well as ex vivo models of mouse and human lung tissues. To define the functional roles of dopamine signaling we will leverage both cell-specific conditional genetic models as well as well-characterized small molecule inhibitors and dopamine agonists in these systems. Together our studies will define the cellular sources and regulatory systems that control dopamine bioavailability during normal lung repair, and will delineate how this repair system fails in human PF. These studies may reveal new therapeutic approaches to promote fibrosis resolution and lung repair.

项目摘要 肺纤维化（PF）仍然是一个重大的医疗负担，治疗选择不令人满意 这种情况无法扭转确定的疾病。我们先前的工作已经确定成纤维细胞中的YAP/TAZ激活是 病理反馈回路的主要特征，该循环传播PF的进展。在先前的资金周期中， 我们将多巴胺D1受体（D1R）激动剂鉴定为在肺中有选择地使YAP/TAZ灭活的策略 成纤维细胞，导致小鼠实验性肺纤维化的加速分辨率部分通过切换 肺成纤维细胞从基质沉积到基质降解状态。而且，我们证明了 PF的个体表现出DOPA脱羧酶（DDC）的表达不足，该酶是催化的酶 将L-DOPA转化为生物活性多巴胺。这些研究使我们提出了恢复 肺中内源性局部多巴胺水平是基质降解和纤维化的重要触发因素 对于成功修复肺而言至关重要，但在PF中受到损害。我们的初步数据显示 博来霉素损伤后，DDC转录本在年轻小鼠的肺组织中瞬时降低并增加 在纤维化分辨率期间，老年小鼠在DDC中表现出持续的降低，这是平行的 纤维化。此外，从第21天到第42天，小分子抑制DDC酶活性或D1R 年轻小鼠中的博霉素消除了肺纤维化的自发分辨率，证明了至关重要的作用 用于纤维化分辨率中的多巴胺信号传导。此外，我们发现多巴胺在 精确切割的肺切片，并从纤维化肺培养的切片中减少，确认局部合成 肺中多巴胺的基于这些发现，我们建议测试上皮的中心假设 多巴胺合成对于纤维化的分辨率和PF肺中正常多巴胺水平的恢复至关重要 组织可以促进肺的胶原蛋白吸收和修复。我们将以三个目标检验这一假设 涵盖肺纤维化的非分辨小鼠模型以及小鼠和人的离体模型 肺组织。为了定义多巴胺信号的功能作用，我们将利用两个细胞特异性条件 遗传模型以及特征良好的小分子抑制剂和多巴胺激动剂在其中 系统。我们的研究将共同​​定义控制多巴胺的细胞来源和调节系统 正常肺修复期间的生物利用度，并将描述该修复系统在人类PF中的失败。这些 研究可能揭示了促进纤维化分辨率和肺修复的新治疗方法。

项目成果

A multiwell platform for studying stiffness-dependent cell biology.

• DOI: 10.1371/journal.pone.0019929
• 发表时间: 2011
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Mih JD;Sharif AS;Liu F;Marinkovic A;Symer MM;Tschumperlin DJ
• 通讯作者: Tschumperlin DJ

GPCR-mediated YAP/TAZ inactivation in fibroblasts via EPAC1/2, RAP2C, and MAP4K7.

• DOI: 10.1002/jcp.30459
• 发表时间: 2021-11
• 期刊: Journal of cellular physiology
• 影响因子: 5.6
• 作者: Choi KM;Haak AJ;Diaz Espinosa AM;Cummins KA;Link PA;Aravamudhan A;Wood DK;Tschumperlin DJ
• 通讯作者: Tschumperlin DJ

Lysophosphatidic acid stimulates epidermal growth factor-family ectodomain shedding and paracrine signaling from human lung fibroblasts.

• DOI: 10.1111/j.1524-475x.2010.00655.x
• 发表时间: 2011-03
• 期刊: Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society
• 作者: Shiomi T;Boudreault F;Padem N;Higashiyama S;Drazen JM;Tschumperlin DJ
• 通讯作者: Tschumperlin DJ

Matrix, mesenchyme, and mechanotransduction.

• DOI: 10.1513/annalsats.201407-320mg
• 发表时间: 2015-04
• 期刊: Annals of the American Thoracic Society
• 影响因子: 8.3
• 作者: D. Tschumperlin

Feedback amplification of fibrosis through matrix stiffening and COX-2 suppression.

• DOI: 10.1083/jcb.201004082
• 发表时间: 2010-08-23
• 期刊: The Journal of cell biology
• 作者: Liu F;Mih JD;Shea BS;Kho AT;Sharif AS;Tager AM;Tschumperlin DJ
• 通讯作者: Tschumperlin DJ