Role of Epac1 in the Pathogenesis of Pulmonary Fibrosis

Epac1在肺纤维化发病机制中的作用

• 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
• 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

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.

项目摘要 该提案的总体目的是描述CAMP直接激活的交换蛋白的作用 （EPAC1）特发性肺纤维化（IPF）疾病。 IPF的特征是进行性疤痕和纤维化 导致威胁生命的并发症（例如呼吸衰竭）的肺部。不幸的是，没有 治愈IPF，我们的衰老世界人口需要绝大多数新的治疗目标和策略 用于治疗这种致命疾病的患者。 大多数纤维化研究都集中在增加营地，以抑制成纤维细胞增殖和通过 但是，PKA途径既不是表达的调节，也不是EPAC1对IPF的贡献 病理生理过程的定义很好。那是有趣的 在IPF中独立于PKA来调节EPAC酶的作用。最近，化合物AM-001具有 被鉴定为一种新型和潜在的EPAC1药物抑制剂。 AM-001有选择地 抑制EPAC1催化活性并显示心脏保护特性。这样的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。我们的初步 研究表明，IPF患者的肺组织中EPAC1的表达显着增加，IPF解散了 与对照组相比，成纤维细胞和博来霉素（BLM）挑战小鼠。此外，EPAC1缺乏小鼠 受到BLM诱导肺损伤和纤维化的保护。此外，EPAC1的敲低和抑制 通过AM-001减弱正常和IPF成纤维细胞增殖和促纤维化标记的表达，TGFβ 和白介素6（IL-6）。另外，AM-001显着降低了BLM诱导的PF体内肺纤维化 鼠标模型。 RNA测序数据分析显示了IPF的促纤维基因标志的关键组成部分 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是重要的调节剂 成纤维细胞在PF中的病理状态，并且EPAC1可以作为AM-001的潜在治疗靶标 用于PF疾病。在此提案中，我们将通过测试以下特定目的来扩展这些初步发现： 特定目的1：用PF定义人类和小鼠的EPAC1的表达模式。 特定目标2：定义成纤维细胞激活中EPAC1功能的机理。 特定目的3：评估新型EPAC1特异性抑制剂AM-001在小鼠模型中的有效性 肺纤维化。总的来说，拟议的研究将有助于提高我们对EPAC1在肺中的作用的理解 与老化的PF发病机理相关的重塑，可能导致鉴定出新的潜在靶标 阻止这种致命疾病的进展。