Correcting Pathogenic TGF beta Activity in the Airway

纠正气道中的致病性 TGF β 活性

基本信息

批准号：
10189898
负责人：
Agnieszka Swiatecka-Urban
金额：
$ 44.66万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-02-01 至 2022-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10189898
关键词：
3&apos Untranslated Regions Affect Air Pollutants Apical Binding Binding Sites Caucasians Cause of Death Cells Chloride Channels Chloride Ion Chronic Obstructive Airway Disease Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Data Defect Dehydration Delta F508 mutation Disabled Persons Disease Epithelial Cells Epitopes FDA approved Financial compensation Gene Mutation Genes Genetic Polymorphism Genetic Translation Health Care Costs Homeostasis Intervention Ion Channel Ion Transport Lead Learning Lung Lung diseases Mediating Mediator of activation protein Messenger RNA MicroRNAs Molecular Movement Mutation Pathogenicity Patients Pharmaceutical Preparations Phenotype Potassium Channel Probability Proteins Pulmonary Cystic Fibrosis Pulmonary Function Test/Forced Expiratory Volume 1 Recovery Regulation Regulator Genes Repression Residual state Resistance Respiratory physiology Response Elements Role Severities Signal Transduction Site Sodium Chloride Structure of parenchyma of lung Testing Therapeutic Transforming Growth Factor beta Transforming Growth Factors Translations Treatment Efficacy Up-Regulation VX-770 VX-809 Water airway epithelium airway surface liquid bronchial epithelium cigarette smoke cystic fibrosis patients design functional restoration genetic variant improved improved outcome inhibitor/antagonist injured airway mRNA Stability mutant novel novel strategies novel therapeutic intervention novel therapeutics nucleocytoplasmic transport preservation prevent pulmonary function targeted treatment

项目摘要

ABSTRACT Homeostasis of the airway surface liquid (ASL) in lung is critically dependent on chloride ion (Cl-) transport, mediated by the apical channel, CFTR (cystic fibrosis transmembrane conductance regulator). CFTR gene mutations associated with the most common autosomal recessive disease in Caucasians, cystic fibrosis (CF) result in loss of CFTR function and severe impairment of the ASL volume regulation. Deletion of F508 (F508del) in the CFTR gene is present in 90% of CF patients; it blocks CFTR biosynthetic processing, reduces CFTR Cl- channel function, and decreases CFTR mRNA stability and translation. It is estimated that the classic severe CF phenotype develops when the CFTR channel function is less than 1% of normal and at least 10% function is needed to alleviate the severe phenotype. Therapies targeting the basic molecular defects in CF demonstrate potential but are insufficient for most patients. While the FDA-approved drug VX-770 potentiates CFTR channel “open” probabilities and improves lung function for less than 10% of patients with the rare mutation G551D, the combined use of VX-770 and a corrector VX-809 that rescues the folding and processing defect was only marginally effective for F508del patients. CF patients with more severe lung disease were entirely resistant to such therapy. Transforming growth factor (TGF)-β1 contributes to resistance of corrector therapy by blocking CFTR translation and represses ancillary ion channels critical to residual ASL homeostasis in CF, namely BK (a Ca+2-activated K+ channel) and ANO1 (a Ca+2-activated Cl- channel). High TGF-β1 levels are seen in 40% of F508del homozygous patients due to polymorphisms in the TGF-β1 gene. Air pollutants, including cigarette smoke also increase TGF-β1 levels. The concomitant upregulation of TGF-β1 in turn increases the severity of CF lung disease in F508del homozygous patients and presents a major block to therapies aimed at F508del-CFTR rescue. We propose that targeting the pathogenic TGF-β1 activity would improve the residual ASL volume homeostasis in CF by restoring the function of BK and ANO1 channels. It would also increase the efficacy of therapy for F508del CF patients by restoring the diminished 508del-CFTR translation and allowing increased activity of the mutant CFTR channel function. Thus, defining TGF-β1 mediators in lung tissue presents an opportunity to design novel drugs eliminating the pathogenic TGF-β1 activity in CF patients. Our central hypothesis is that the targeted reduction of TGF-β1 activity will ameliorate negative effects on CFTR, BK, and ANO1 channels and ASL homeostasis, and provide a novel approach to treat F508del CF patients. Our specific aims are to test the hypothesis: (1) that Dab2 is a TGF-β1 adaptor that inhibits the residual ASL homeostasis in CF bronchial epithelium by directing nuclear transport of Smad3 to repress the ancillary channels, BK and ANO1; (2) that the Dab2-Smad3 interactions upregulates microRNAs that repress the ancillary channels and block corrector-mediated rescue of F508del-CFTR; and (3) that a Dab2-Smad3 interaction is required for the pathogenic activity of TGF-β1 in CF bronchial epithelium.

抽象的肺中气道表面液体（ASL）的稳态严重取决于氯离子（Cl-）转运，由顶端通道CFTR（囊性纤维化跨膜电导调节剂）介导。 CFTR基因与高加索人最常见的常染色体隐性疾病相关的突变，囊性纤维化（CF）导致CFTR功能的丧失和ASL体积调节严重损害。删除F508 CFTR基因中的（F508DEL）存在于90％的CF患者中；它阻止CFTR生物合成处理，减少 CFTR CL-通道功能，并降低CFTR mRNA稳定性和翻译。据估计经典当CFTR通道函数小于正常和至少10％时，严重的CF表型开发需要功能来减轻严重的表型。针对CF基本分子缺损的疗法具有潜力，但对于大多数患者来说不足。而FDA批准的药物VX-770潜力 CFTR渠道“开放”可能性，并改善少于10％的罕见患者的肺功能突变G551D，VX-770和校正vx-809的综合使用响应折叠和处理缺陷仅对F508DEL患者有略有有效的效果。 CF患者患有更严重的肺病患者转化生长因子（TGF）-β1有助于校正器的抗性通过阻止CFTR翻译并反映对残留ASL稳态至关重要的辅助离子通道的治疗在CF中，即BK（一个Ca+2个激活的K+通道）和ANO1（Ca+2个激活的Cl-通道）。高TGF-β1水平由于TGF-β1基因的多态性，在40％的F508DEL纯合患者中看到了40％。空气污染物，包括香烟烟雾还增加了TGF-β1水平。 TGF-β1的同时上调增加了F508DEL纯合患者的CF肺病的严重程度，并为针对F508DEL-CFTR救援的疗法。我们提出，针对致病性TGF-β1活性将通过恢复BK和ANO1通道的功能，改善CF中的残留ASL体积稳态。它还将通过恢复降低的508DEL-CFTR来提高F508DEL CF患者的治疗效率翻译并允许突变CFTR通道功能的活动增加。这是定义TGF-β1 肺组织中的介质提供了设计新型药物消除致病性TGF-β1的机会 CF患者的活性。我们的中心假设是，TGF-β1活性的靶向降低将改善对CFTR，BK和ANO1通道和ASL稳态的负面影响，并提供了一种新颖的方法治疗F508DEL CF患者。我们的具体目的是检验假设：（1）DAB2是TGF-β1适配器通过将SMAD3的核转运引向CF支气管上皮的残留ASL稳态稳态抑制辅助频道，BK和ANO1；（2）DAB2-SMAD3相互作用上调microRNA 这反映了辅助通道和阻塞校正器介导的F508DEL-CFTR的营救；（3） DAB2-SMAD3相互作用是TGF-β1在CF支气管上皮细胞中的致病活性所必需的。