Role of Epac1 in the Pathogenesis of Pulmonary Fibrosis

Epac1在肺纤维化发病机制中的作用

基本信息

批准号：
10594558
负责人：
Lahouaria HADRI
金额：
$ 42.25万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10594558
关键词：
Adenoviruses Affect Aging Attenuated Biological Assay Biological Process Biopsy Bleomycin Cardiovascular Diseases Cell Adhesion Cells Chest Chronic Cicatrix Cyclic AMP Cyclic AMP-Dependent Protein Kinases Data Data Analyses Development Disease Enzymes Exposure to FGFR1 gene FOXO3A gene Fibroblasts Fibrosis Foundations Goals Growth Guanosine Triphosphate Phosphohydrolases Histology Human Immune response Immunofluorescence Immunologic In Vitro Inflammation Inflammatory Interleukin-6 Knockout Mice Knowledge Life Lung Lung diseases MADH2 gene Malignant Neoplasms Measurement Messenger RNA Molecular Mus Pathogenesis Pathologic Pathway interactions Patients Pattern Phenotype Phosphorylation Population Predisposition Process Profibrotic signal Proliferating Property Proteins Proto-Oncogene Proteins c-akt Pulmonary Fibrosis Quantitative Reverse Transcriptase PCR Regulation Research Respiratory Failure Respiratory physiology Role STAT3 gene Scanning Severity of illness Shortness of Breath Signal Pathway Signal Transduction Structure of parenchyma of lung Tenascin Testing Therapeutic Effect Time Tissue Sample Transforming Growth Factor beta Western Blotting aged cardioprotection cell growth comparison control coronary fibrosis cytokine effectiveness evaluation experimental study fibrotic lung gain of function genetic signature idiopathic pulmonary fibrosis in vivo indium-bleomycin inhibitor knock-down loss of function lung injury microCT migration mortality mouse model myocardial injury new therapeutic target novel novel strategies novel therapeutics overexpression pharmacologic promoter pulmonary function ras Guanine Nucleotide Exchange Factors response sensor small hairpin RNA therapeutic target tool transcription factor transcriptome sequencing

项目摘要

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.

项目概要该提案的总体目标是描述由 cAMP 直接激活的交换蛋白的作用 (Epac1) 在特发性肺纤维化 (IPF) 疾病中的特征是进行性疤痕形成和纤维化。不幸的是，没有这种情况会导致呼吸衰竭等危及生命的并发症。治愈 IPF 以及世界人口老龄化需要绝大多数新的治疗靶点和策略用于治疗患有这种致命疾病的患者。大多数纤维化研究都集中在增加 cAMP 以通过以下方式抑制成纤维细胞增殖和分化：然而，PKA 通路既没有表达调节，也没有 Epac1 对 IPF 的贡献。因此，通过具体指导研究是很有趣的。最近，化合物 AM-001 在 IPF 中独立于 PKA 调节 Epac 酶的作用。 AM-001选择性抑制 Epac1 催化活性并显示出心脏保护特性，这些发现反映了评估的必要性。 Epac1 及其特异性抑制剂 AM-001 在肺纤维化 (PF) 疾病中的关键作用。研究表明，IPF患者肺组织中Epac1表达显着升高，IPF患病与对照组相比，成纤维细胞和博来霉素 (BLM) 攻击的小鼠。此外，Epac1 缺陷小鼠可以防止 BLM 诱导的肺损伤和纤维化此外，Epac1 的敲低和抑制。 AM-001 减弱正常和 IPF 成纤维细胞增殖以及促纤维化标志物 TGFβ 的表达此外，AM-001 还可显着降低 BLM 诱导的 PF 体内肺纤维化。 RNA 测序数据分析显示了 IPF 促纤维化基因特征的关键组成部分。在 AM-001 处理的 NHLF 细胞中，基于这些数据，我们的总体假设是 Epac1 是一个重要的调节因子。 PF 中成纤维细胞的病理状态以及 Epac1 可以作为 AM-001 的潜在治疗靶点在本提案中，我们将通过测试以下具体目标来扩展这些初步发现：具体目标 1：确定 Epac1 在患有 PF 的人类和小鼠中的表达模式。具体目标 2：明确 Epac1 在成纤维细胞激活中的功能机制。具体目标 3：评估新型 Epac1 特异性抑制剂 AM-001 在小鼠模型中的有效性总的来说，拟议的研究将有助于增加我们对 Epac1 在肺中作用的理解。与老年 PF 发病机制相关的重塑，可能导致新的潜在靶点的识别阻止这种致命疾病的进展。