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，可逆的障碍物外，某些哮喘患者还出现固定的气道阻塞作为改建。越来越多的文献表明，被激活和寿命长的气道和实质嗜酸性粒细胞（EOS）诱导支气管成纤维细胞，平滑肌和肌上皮细胞以增强的胶原蛋白和细胞外基质蛋白的产生为顶点。从气道中消除嗜酸性粒细胞时减少了重塑，表明抑制 EOS衍生的TGFPL或在细胞靶标中其纤维化信号传导的阻塞可能具有治疗益处。最近，我们确定了PIN1，肽基 - 蛋白酶异构酶（PPIASE）是TGF的参与者（生产31 和信号。 PIN1与环磷脂A和FKBP有关，是唯一已知的结合的真核生物酶并催化磷酸苯胺或磷酸硫氰胺 - 丙啉肽的顺式传播异构化债券。靶蛋白的异构化改变其构象，功能或稳定性。 PIN1最近有与GM-CSF mRNA代谢有关，表明在细胞因子的调节中起着更广泛的作用活化的EOS生物发生。现在，我们提供证据表明PIN1调节EOS和EOS的TGFPL表达成纤维细胞中的TGFPL信号传导。 EOS中PINVS PPIASE活性的特异性封锁降低了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，品脱与已涉及的多种蛋白质结合调节TGFPL mRNA衰减或基因表达。 PIN1抑制减少了胶原mRNA的积累在支气道气道中，衍生的成纤维细胞在体外和气道中暴露于TGFPL，支气管肺泡在体内治疗的哮喘过敏大鼠模型的灌洗（BAL）细胞和肺实质。因此，我们假设PIN1是一个关键的信号中间体，在生产和动作中起关键作用哮喘肺中的TGFPL。因此，我们建议：1。确定如何调节PIN1异构酶活性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，并确定PIN1是否阻断可以改变慢性过敏原暴露动物模型中的气道重塑。在总体中，这些研究将阐明PIN1在哮喘发病机理和气道重塑中的作用和功能。