Role of Epac1 in the Pathogenesis of Pulmonary Fibrosis

项目总结 这项提案的总体目标是描述cAMP直接激活的交换蛋白的作用。 (Epac1)在特发性肺纤维化(IPF)疾病中的表达。IPF的特点是进行性瘢痕形成和纤维化。 导致呼吸衰竭等危及生命的并发症的肺部疾病。不幸的是，没有 治愈IPF和我们老龄化的世界人口需要绝大多数新的治疗目标和战略 治疗这种致命疾病的病人。 大多数关于纤维化的研究都集中在cAMP的增加，通过 然而，PKA通路既不调节Epac1的表达，也不调节Epac1对IPF的贡献 病理生理过程被很好地定义。因此，通过具体的方式来指导研究是很有趣的 在IPF中独立于PKA调节EPAC酶的作用。最近，化合物AM-001已经 已被鉴定为一种新的、有效的Epac1药理抑制剂。AM-001选择性 抑制Epac1的催化活性并显示出心脏保护特性。这些调查结果反映了评估的必要性 Epac1及其特异性抑制物AM-001在肺纤维化疾病中的关键作用我们的预赛 研究表明，IPF患者肺组织中Epac1的表达显著增加，IPF患者肺组织中Epac1的表达明显增加。 成纤维细胞，以及博莱霉素(BLM)攻击的小鼠与对照组进行比较。此外，Epac1缺陷小鼠 对博莱姆诱导的肺损伤和纤维化具有保护作用。此外，Epac1基因的敲除和抑制 AM-001抑制正常和特发性肺纤维化成纤维细胞增殖及促纤维化标志物转化生长因子β的表达 白介素6(IL-6)。此外，AM-001还能显著减轻博莱姆所致肺纤维化大鼠的体内肺纤维化。 老鼠模型。RNA测序数据分析显示IPF促纤维化基因签名的关键成分 在AM-001处理的NHLF细胞中。根据这些数据，我们的总体假设是Epac1是一个重要的调节因子 成纤维细胞在PF中的病理状态及Epac1可作为AM-001潜在的治疗靶点 用于治疗肺泡炎。在这项提案中，我们将通过测试以下具体目标来扩展这些初步调查结果： 具体目的1：明确Epac1在人和PF小鼠中的表达模式。 特异性目的2：明确Epac1在成纤维细胞活化中的作用机制。 具体目的3：评价新型Epac1特异性抑制剂AM-001在小鼠卵巢癌模型中的疗效。 肺纤维化。总的来说，建议的研究将有助于加深我们对Epac1在肺部的作用的了解。 重构与AGE-PF的发病机制相关，并可能导致识别新的潜在靶点 阻止这种致命疾病的发展。

PROJECT SUMMARY The overall objective of this proposal is to delineate the role of the exchange protein directly activated by cAMP (Epac1) in idiopathic pulmonary fibrosis (IPF) disease. IPF is characterized by progressive scarring and fibrosis of the lungs that results in life-threatening complications such as respiratory failure. Unfortunately, there is no cure for IPF, and our aging world population requires a vast majority of new therapeutic targets and strategies for treating patients with this fatal disease. Most studies in fibrosis focused on increased cAMP to inhibit fibroblast proliferation and differentiation through the PKA pathway, however, neither the regulation of the expression nor the contribution of Epac1 to IPF pathophysiological processes are well defined. Thus, it is interesting to direct the research by specifically regulating the actions of the Epac enzymes independently of PKA in IPF. Recently, the compound AM-001 has been identified and characterized as a novel and potent Epac1 pharmacological inhibitor. AM-001 selectively inhibits Epac1 catalytic activity and displays cardioprotective properties. Such findings reflect the need to assess the key role of Epac1 and its specific inhibitor AM-001 in pulmonary fibrosis (PF) disease. Our preliminary studies show that the expression of Epac1 is significantly increased in lung tissue from IPF patients, IPF diseased fibroblasts, and bleomycin (BLM)-challenged mice compared to controls. Furthermore, Epac1 deficiency mice are protected against BLM induced-lung injury and fibrosis. Furthermore, the knockdown and inhibition of Epac1 by AM-001 attenuate normal and IPF fibroblasts proliferation and the expression of pro-fibrotic markers, TGFβ and interleukin-6 (IL-6). In addition, AM-001 significantly decreases lung fibrosis in vivo in the BLM-induced PF mouse model. RNA sequencing data analyses show key components of the pro-fibrotic genes signature of IPF in AM-001-treated NHLF cells. Based on these data our overall hypothesis is that Epac1 is an important regulator of the fibroblast's pathological state in PF and that Epac1 can serve as a potential therapeutic target by AM-001 for PF disease. In this proposal, we will extend these preliminary findings by testing the following specific aims: Specific Aim 1: To define the expression pattern of Epac1 in humans and mice with PF. Specific Aim 2: To define the mechanisms of Epac1 function in fibroblast activation. Specific Aim 3: To evaluate the effectiveness of a novel Epac1-specific inhibitor AM-001 in a mouse model of pulmonary fibrosis. Collectively, the proposed studies will help increase our understanding of Epac1 role in lung remodeling associated with aged-PF pathogenesis and may lead to the identification of new potential targets to block the progression of this deadly disease.