Control of fibrosing lung disease by p53-miR-34a-targeted therapeutics

通过 p53-miR-34a 靶向疗法控制纤维化肺疾病

• 批准号: 9157281
• 负责人: Sreerama Shetty
• 金额: $ 39.18万
• 依托单位: UNIVERSITY OF TEXAS HLTH CTR AT TYLER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9157281
• 关键词: Address; Affect; Alveolar; Amino Acids; Anti-Inflammatory Agents; Anti-inflammatory; Apoptosis; Architecture; Biochemical; Bleomycin; Cells; Cicatrix; Cytoskeletal Modeling; Data; Deposition; Development; Diagnosis; Disease; Epithelial; Epithelial Cells; Etiology; Europe; Extracellular Matrix; Fibroblasts; Fibrosis; Functional disorder; Generations; Hamman-Rich syndrome; Histologic; Human; Imaging Techniques; Interstitial Lung Diseases; Intervention; Knockout Mice; Lung; Lung diseases; Mediating; MicroRNAs; Modeling; Molecular; Mus; Myofibroblast; Pathogenesis; Pathway interactions; Patients; Peptides; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Platelet-Derived Growth Factor; Population; Process; Production; Proliferating; Property; Protein p53; Proteins; Pulmonary Fibrosis; Regulation; Reporting; Resistance; Respiratory physiology; Role; Signal Transduction; Staging; Structure of parenchyma of lung; TP53 gene; Tertiary Protein Structure; Testing; Time; Tissues; Tumor Suppressor Proteins; Ubiquitination; Work; attenuation; base; biochemical model; caveolin 1; feeding; improved; improved outcome; interstitial; lung injury; migration; molecular imaging; novel; outcome forecast; prevent; repaired; response; restoration; scaffold; targeted treatment

Idiopathic pulmonary fibrosis (IPF) is the most common interstitial lung disease and is fatal. It affects 500,000 people in the USA and Europe per year with a median survival of <2-3 years after diagnosis. Its pathogenesis is poorly understood and there is no cure. A hallmark of IPF is the formation of fibroblastic foci, consisting of large numbers of fibrotic lung fibroblasts (fLFs) and interstitial fibrosis with architectural distortion. The population of fLFs in the lungs of patients with IPF includes myofibroblasts and activated highly proliferative, migratory fibroblasts that deposit excessive matrix proteins. Our preliminary data demonstrate that basal expression of p53 and microRNA-34a (miR-34a), are markedly reduced, while expression of platelet derived growth factor- (PDGFR- is significantly increased in fLFs from IPF lungs compared with control fibroblasts extracted from histologically “normal” lungs (nLFs). This also occurs in mice with bleomycin (BLM)-induced pulmonary fibrosis (PF). In addition, p53 and miR-34a act through an autoregulatory feed-forward loop, which in turn suppresses PDGFR-. Our working hypothesis is that p53 and miR-34a are reduced in fLFs, which induce pro-fibrogenic responses in fLFs that are critical to the pathogenesis of PF. We infer that restoration of p53 and miR-34a in fLFs will mitigate PF. Our objective is to elucidate the mechanism by which reduced p53 and miR-34a expression in fLFs promote PF. We will also determine if targeting of this pathway suppresses pro-fibrogenic properties of fLFs and thereby reverses established PF. The peptide CSP7 (FTTFTVT) will be used to target this pathway. We have previously shown that the p53-targeting with CSP7 blocks alveolar epithelial apoptosis and prevents development of BLM-induced PF. However, the effects of CSP7 on fLFs are unknown, as is how targeting of the p53/miR-34a pathway in these cells resolves established PF. These represent potentially critical gaps in our understanding of the pathogenesis of PF and will be addressed in this project. Our Aims are: 1) To elucidate the role of p53 in pro-fibrogenic responses in fLFs. We will define mechanism by which p53 regulates pro-fibrogenic responses of fLFs and test the ability CSP7 to block these responses in fLFs from patients with IPF and from mice with BLM- or TGF--induced PF. 2) To determine how miR-34a regulates p53-mediated pro-fibrogenic responses in fLFs. We will define the role of altered miR-34a expression in the control of these responses in fLFs and test whether the same interventions reverse the miR- 34a-mediated responses. 3) To elucidate the signaling mechanisms involved in p53- and miR-34a-mediated regulation of pro-fibrogenic responses in LFs. State of the art modeling, biochemical, molecular and imaging- based approaches will be used to accomplish the aims. This project will have a major impact on the field by determining, for the first time, how p53 and miR-34a regulate pro-fibrotic responses including changes in PDGFR- in fLFs and PF. The work may also define novel, p53/miR-34a-targeted approaches to reverse these effects and improve outcomes for patients with IPF.

特发性肺纤维化（IPF）是最常见的间质性肺疾病，并且是致命的。影响50万 美国和欧洲每年诊断后中位生存期<2-3年。其发病机制 人们对此知之甚少，并且无法治愈。 IPF 的一个标志是成纤维细胞灶的形成，其中包括 大量纤维化肺成纤维细胞（fLF）和伴有结构扭曲的间质纤维化。这 IPF 患者肺部的 fLF 群体包括肌成纤维细胞和激活的高度增殖、 沉积过量基质蛋白的迁移性成纤维细胞。我们的初步数据表明，基础 p53 和 microRNA-34a (miR-34a) 的表达显着降低，而血小板衍生的表达 与对照成纤维细胞相比，IPF 肺部的 fLF 中生长因子-（PDGFR- 显着增加） 从组织学上“正常”的肺（nLF）中提取。这种情况也发生在博莱霉素 (BLM) 诱导的小鼠身上 肺纤维化（PF）。此外，p53 和 miR-34a 通过自动调节前馈环发挥作用， 进而抑制 PDGFR-。我们的工作假设是 p53 和 miR-34a 在 fLF 中减少，这 在 fLF 中诱导促纤维化反应，这对 PF 的发病机制至关重要。我们推断恢复 fLF 中的 p53 和 miR-34a 将减轻 PF。我们的目标是阐明减少 p53 的机制 fLF 中 miR-34a 的表达可促进 PF。我们还将确定该通路的靶向是否会抑制 fLF 的促纤维化特性，从而逆转已建立的 PF。肽 CSP7 (FTTFTVT) 将是 用于靶向该途径。我们之前已经证明，以 CSP7 为靶标的 p53 会阻断肺泡 上皮细胞凋亡并防止 BLM 诱导的 PF 的发展。然而，CSP7 对 fLF 的影响是 尚不清楚，这些细胞中的 p53/miR-34a 通路的靶向如何解决已建立的 PF 也是未知的。这些 代表了我们对 PF 发病机制的理解中潜在的关键差距，并将在本文中得到解决 项目。我们的目标是：1) 阐明 p53 在 fLF 促纤维化反应中的作用。我们将定义 p53 调节 fLF 促纤维化反应的机制，并测试 CSP7 阻断这些反应的能力 IPF 患者和 BLM 或 TGF-β 诱导的 PF 小鼠的 fLF 反应。 2）确定如何 miR-34a 调节 fLF 中 p53 介导的促纤维化反应。我们将定义改变的 miR-34a 的作用 在 fLF 中控制这些反应的表达并测试相同的干预措施是否逆转 miR- 34a介导的反应。 3) 阐明p53和miR-34a介导的信号传导机制 LF 中促纤维化反应的调节。最先进的建模、生化、分子和成像- 将使用基于基础的方法来实现目标。该项目将对该领域产生重大影响 首次确定 p53 和 miR-34a 如何调节促纤维化反应，包括 fLF 和 PF 中的 PDGFR-。这项工作还可能定义新颖的 p53/miR-34a 靶向方法来逆转这些 效果并改善 IPF 患者的预后。