Regulation of TGF-B1 Production and Signaling by Pin-1

Pin-1 对 TGF-B1 产生和信号传导的调节

基本信息

批准号：
7843281
负责人：
James S Malter
金额：
$ 34.52万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-02-01 至 2013-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7843281
关键词：
Affect Allergens Allergic Animal Model Asthma Binding Biogenesis Bronchoalveolar Lavage Cell Cycle Progression Cells Chronic Collagen Complex Cyclophilin A Deposition Elasticity Enzymes Event Extracellular Matrix Proteins Fibroblasts Fibrosis Gene Expression Granulocyte-Macrophage Colony-Stimulating Factor Hyperplasia Immunoprecipitation In Vitro Isomerase Life Literature Lung Messenger RNA Metabolism Modeling Molecular Conformation Myoepithelial cell Nerve Degeneration Obstruction Participant Pathogenesis Pathway interactions Patients Peptides Peptidylprolyl Isomerase Phosphoserine Phosphothreonine Pin1 protein Play Process Production Proline Proteins Rattus Regulation Respiratory physiology Role Signal Transduction Smooth Muscle Structure of parenchyma of lung Tacrolimus Binding Proteins Therapeutic Transgenic Mice airway inflammation airway obstruction airway remodeling asthmatic airway cellular targeting cis trans isomerization cytokine eosinophil experience flexibility in vivo mRNA Decay mRNA Stability parvulin protein expression receptor-mediated signaling tumorigenesis

项目摘要

In addition to intermittent, reversible obstruction, some asthmatics, develop fixed airway obstruction known as remodeling. A growing literature suggests thatTGFpt produced by activated and long-lived airway and parenchymal eosinophils (EOS) induces hyperplasia of bronchiolar fibroblasts, smooth muscle and myoepithelial cells culminating with enhanced production of collagens and extracellular matrix proteins. Remodeling is reduced when eosinophils are eliminated from the airways suggesting that suppression of EOS derived TGFpl or blockade of its profibrotic signaling in cellular targets could have therapeutic benefits. Recently, we have identified Pin1, a peptidyl-prolyl isomerase (PPIase) as a participant in TGF(31 production and signaling. Pin1 is related to cyclophilin A and FKBP and is the only known eukaryotic enzyme that binds to and catalyzes the cis-trans isomerization of phosphoserine-proline or phosphothreonine-proline peptide bonds. The isomerization of target proteins alters their conformation, function or stability. Pin1 has recently been implicated in GM-CSF mRNA metabolism, suggesting a broader role in the regulation of cytokine biogenesis by activated EOS. We now present evidence that Pin1 regulates TGFpl expression by EOS and TGFpl signaling in fibroblasts. Specific blockade of PinVs PPIase activity in EOS reduced TGFpl mRNA stability, causing steady state mRNA levels and protein expression to significantly decline. Immunoprecipitation studies revealed that Pint binds to multiple proteins that have been implicated in the regulation of TGFpl mRNA decay or gene expression. Pin1 inhibition reduced collagen mRNA accumulation in bronchial airway derived fibroblasts exposed to TGFpl in vitro as well as in the airways, bronchoalveolar lavage (BAL) cells and lung parenchyma of allergic rat models of asthma treated in vivo. Therefore, we hypothesize that Pin1 is a critical, signaling intermediate that plays a key role in the production and action of TGFpl in the asthmatic lung. We therefore propose to: 1. Identify how Pin1 isomerase activity is regulated after eosinophil activation, 2. Determine how Pin1 protein targets HuR and AUF1 control TGFpl mRNA decay, 3. Determine how Pin1 regulates TGFpl signaling in fibroblasts and 4, Determine if Pin1 blockade can alter airway remodeling in animal models of chronic allergen exposure. In aggregate these studies will clarify the role and function of Pin1 in asthma pathogenesis and airway remodeling.

除了间歇性、可逆性阻塞外，一些哮喘患者还会出现已知的固定气道阻塞作为改造。越来越多的文献表明，TGFpt 由活化且长寿命的气道产生，实质嗜酸性粒细胞（EOS）诱导细支气管成纤维细胞、平滑肌和肌上皮细胞最终增强胶原蛋白和细胞外基质蛋白的产生。当嗜酸性粒细胞从气道中消除时，重塑就会减少，这表明抑制 EOS 衍生的 TGFpl 或阻断其在细胞靶标中的促纤维化信号传导可能具有治疗益处。最近，我们发现 Pin1，一种肽基脯氨酰异构酶 (PPIase)，作为 TGF(31 生产) 的参与者和信号。 Pin1 与亲环蛋白 A 和 FKBP 相关，是唯一已知的结合亲环蛋白 A 和 FKBP 的真核酶形成并催化磷酸丝氨酸-脯氨酸或磷酸苏氨酸-脯氨酸肽的顺反异构化债券。靶蛋白的异构化改变了它们的构象、功能或稳定性。 Pin1 最近与 GM-CSF mRNA 代谢有关，表明在细胞因子的调节中具有更广泛的作用由激活的 EOS 进行生物发生。我们现在提供的证据表明 Pin1 通过 EOS 调节 TGFpl 表达成纤维细胞中的TGFβ1信号传导。 EOS中PinVs PPIase活性的特异性阻断减少了TGFpl mRNA 稳定性，导致稳态 mRNA 水平和蛋白质表达显着下降。免疫沉淀研究表明，Pint 与多种蛋白质结合，这些蛋白质与 TGFβ1 mRNA衰减或基因表达的调节。 Pin1 抑制减少胶原 mRNA 积累在体外暴露于 TGFpl 的支气管气道来源的成纤维细胞以及气道、支气管肺泡中体内治疗的过敏性哮喘大鼠模型的灌洗（BAL）细胞和肺实质。因此，我们假设 Pin1 是一种关键的信号中间体，在哮喘肺中的TGFpl。因此，我们建议： 1. 确定 Pin1 异构酶活性是如何调节的嗜酸性粒细胞激活后，2.确定Pin1蛋白如何靶向HuR和AUF1控制TGFpl mRNA 衰减，3. 确定 Pin1 如何调节成纤维细胞中的 TGFpl 信号传导，4. 确定 Pin1 是否阻断可以改变慢性过敏原暴露的动物模型中的气道重塑。总的来说，这些研究将阐明Pin1在哮喘发病机制和气道重塑中的作用和功能。