Inhibitors of Epithelial TGFB signaling and EMT/Sheppard

项目总结(见说明)： TGFB1信号在特发性肺纤维化(IPF)的病理生物学中是重要的，但在许多对正常内环境稳定至关重要的生理过程中也是关键的。针对TGFB1信号减弱的抗纤维化药物毒性较小，但如果专注于肝纤维化特异性信号转导方面，则仍有效。到目前为止，在患者中实现这一目标的方法仍然难以捉摸。最近对上皮间质转化(EMT)的研究表明，EMT在实验性肺纤维化过程中在体内发生，并在纤维化过程中起重要作用。EMT被发现严重依赖于整合素a3B1依赖的酪氨酸磷酸化(PY654)B-连环素和pSmad2转录复合体的产生。这些转录复合体存在于IPF肺提取物中，并定位于这种疾病中积聚的肌成纤维细胞的细胞核，这意味着新的治疗方法可以接近TGFB1信号的相对特异的途径。这项应用的目标是开发能够衰减的新试剂 在小鼠体内的EMT和肺纤维化，这可能是该项目临床阶段药物开发的基础。最近发现的抑制pY654-P-连环蛋白生成的整合素a3B1衍生多肽和促进副-B-连环蛋白周转的小分子将是在小鼠肺纤维化模型中进行抑制剂优化和测试的起点。这两类抑制剂都能在体外阻断EMT。此外，模拟整合素衍生的多肽和/或体外阻断EMT的小分子的高通量细胞筛选将被用于开发额外的先导化合物，用于优化和体内测试。该项目将与用于多肽和小分子优化的药物化学核心密切合作，与人体组织核心密切合作，以研究具有原代人类肺泡上皮细胞的有希望的先导化合物，与纵向队列核心合作，以确定具有机械信息的生物标志物的特征，并与项目2密切合作，该项目将开发用于药物输送的气雾剂系统。我们预计，拟议中的研究将检验抑制EMT是减轻纤维化形成的有效方法这一原则，并为进一步开发IPF的治疗药物提供有前景的候选药物。

PROJECT SUMMARY (See instructions): TGFB1 signaling is prominent in the pathobiology of idiopathic pulmonary fibrosis (IPF) but is also critical in many physiological processes important to normal homeostasis. Anti-fibrotic drugs directed at attenuation of TGFB1 signaling could be expected to be less toxic and yet still effective if focused on aspects of signaling specific to fibrogenesis. An approach to achieve this goal in patients has to date remained elusive. Recent studies of epithelial mesenchymal transition (EMT) indicate EMT develops in vivo during experimental lung fibrosis and is important to the fibrotic process. EMT was found to critically depend on integrin a3B1-dependent generation of transcriptional complexes of a tyrosine phosphorylated (pY654) B-catenin and pSmad2. These transcriptional complexes are present in extracts of IPF lungs and localize to nuclei of accumulating myofibroblasts in this disorder, implying a relatively specific pathway of TGFB1 signaling approachable by novel therapeutics. The goal of this application is to develop new reagents that attenuate EMT and pulmonary fibrosis in vivo in mice that could be the basis for drug development in the clinical phase of the project. Recently identified integrin a3B1-derived peptides that block pY654-P-catenin generation and small molecules that promote turnover of BY-B-catenin will be the starting points for inhibitor optimization and testing in models of murine lung fibrosis. Both of these inhibitor classes block EMT ex vivo. In addition a high throughput cell based screen of small molecules that mimic the integrin-derived peptides and/or block EMT ex vivo will be used to develop additional lead compounds for optimization and in vivo testing. The project will work closely with the medicinal chemistry core for peptide and small molecule optimization, with the human tissue core to examine promising lead compounds with primary human alveolar epithelial cells, with the Longitudinal Cohort Core to charactize mechanistically informative biomarkers, and with Project 2 that will develop an aerosol system for drug delivery. We anticipate that the proposed studies should both test the principle that inhibition of EMT is an effective approach to attenuation of fibrogenesis and provide promising candidates for futher development as therapeutics for IPF.