The Role of Iron In Pulmonary Fibrosis

铁在肺纤维化中的作用

• 批准号: 10838699
• 负责人: LIN LIU
• 金额: $ 1.71万
• 依托单位: OKLAHOMA STATE UNIVERSITY STILLWATER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-23 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10838699
• 关键词: Advanced Development; Area; Asbestos; Bleomycin; Chronic lung disease; Co-Immunoprecipitations; Development; Diagnosis; Disease; Down-Regulation; Etiology; Family; Fibroblasts; Genes; Genetic; Genetic Transcription; Goals; Human; In Vitro; Individual; Inhalation; Iron; Lung; Mediating; Modeling; Molecular; Mus; Mutagenesis; Particulate Matter; Pathogenesis; Pathway Analysis; Pathway interactions; Persons; Process; Protein Kinase Interaction; Pulmonary Fibrosis; Regulation; Ring Finger Domain; Risk Factors; Role; SH3 Domains; Signal Pathway; Testing; Ubiquitination; United States; Up-Regulation; cigarette smoke; effective therapy; gain of function; gene therapy; genetic manipulation; homeodomain; idiopathic pulmonary fibrosis; in vivo; inhibitor; insight; loss of function; member; mouse model; novel; pharmacologic; pre-clinical; solute; ubiquitin-protein ligase

PROJECT SUMMARY Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease with a median survival of 3 years after diagnosis. There are no cures for this disease. Although the etiology is still unclear, one of the major risk factors associated with IPF is inhaled iron-rich particulate matter, such as asbestos and cigarette smoke. However, the specific role of iron in the pathogenesis of IPF is an understudied area. The long-term goal of this project is to elucidate the pathogenesis of IPF and thus to advance the development of effective therapies. The objective of the current application is to understand the contribution of iron to the pathogenesis of IPF and the underlying mechanisms with a focus on the iron-mediated activation of lung fibroblasts. There is evidence showing that iron accumulates in the lung and contributes to lung fibrosis. However, the molecular mechanisms for both processes are unclear. Our preliminary studies have identified the iron export gene solute carrier family 40 member 1 (SLC40A1) as a key gene for iron accumulation in lung fibroblasts. We have also identified two novel iron-regulated genes, SH3 domain-containing ring finger 1 (SH3RF1), an E3 ubiquitin protein ligase, and homeodomain-interacting protein kinase 2 (HIPK2), a cotranscriptional regulator and provided evidence that both genes regulate lung fibroblast activation. Based on our preliminary studies, our overall hypothesis is that an elevated level of iron in lung fibroblasts due to reduced SLC40A1 leads to the activation of lung fibroblasts via the downregulation of SH3RF1 expression and the upregulation of HIPK2 expression. The hypothesis will be tested by a combination of in vitro studies using primary human lung fibroblasts and in vivo strategies in two mouse lung fibrosis models using pharmacological and genetic interventions. Aim I will determine the mechanisms by which iron accumulates in lung fibroblasts via SLC40A1 and the regulation and functional roles of SH3RF1 in iron-mediated lung fibroblast activation using loss- and gain-of-function and mutagenesis approaches. Aim II will define the mechanisms by which iron activates lung fibroblasts via HIPK2 using gene manipulation, coimmunoprecipitation, ubiquitination and molecular signaling pathway analyses. Aim III will determine the in vivo effects of iron and HIPK2 on lung fibrosis in two preclinical mouse models of bleomycin- and asbestos-induced lung fibrosis using fibroblast-specific genetic deletion of Hipk2 and pharmacological inhibitors. The proposed studies will establish the molecular mechanisms of iron accumulation in lung fibroblasts and novel roles for SH3RF1 and HIPK2 in lung fibroblast activation.

项目摘要 特发性肺纤维化（IPF）是一种慢性肺部疾病，中位生存期在诊断后3年。 这种疾病没有治疗方法。尽管病因尚不清楚，但主要风险因素之一 使用IPF是吸入铁的颗粒物，例如石棉和香烟烟雾。但是，具体角色 IPF发病机理中的铁是一个研究的区域。该项目的长期目标是阐明 IPF的发病机理，从而促进有效疗法的发展。电流的目的 应用是了解铁对IPF的发病机理和潜在机制的贡献 侧重于铁介导的肺成纤维细胞的激活。 有证据表明铁在肺部积聚并导致肺纤维化。但是， 这两个过程的分子机制尚不清楚。我们的初步研究已经确定了铁的出口 基因溶质载体家族40成员1（SLC40A1）作为肺成纤维细胞中铁积累的关键基因。我们 还鉴定了两个新型铁调节的基因，即含SH3结构域的环形指1（SH3RF1），一个E3 泛素蛋白连接酶和同源域相互作用蛋白激酶2（HIPK2），共转录调节剂和 提供了两个基因调节肺成纤维细胞激活的证据。根据我们的初步研究 总体假设是，由于SLC40A1降低，肺成纤维细胞的铁水平升高导致 通过下调SH3RF1表达和HIPK2的上调激活肺成纤维细胞 表达。该假设将通过使用原发性人肺的体外研究结合进行检验 使用药理和遗传学的两个小鼠肺纤维化模型中的成纤维细胞和体内策略 干预措施。目的我将确定铁在肺成纤维细胞中通过SLC40A1积聚的机制 以及使用损失和 功能收益和诱变方法。 AIM II将定义铁激活肺的机制 通过基因操纵，共免疫沉淀，泛素化和分子信号传导通过HIPK2通过HIPK2进行成纤维细胞 途径分析。 AIM III将确定铁和Hipk2对两个临床前肺纤维化的体内影响 博来霉素和石棉诱导的肺纤维化的小鼠模型使用成纤维细胞特异性遗传缺失的遗传缺失 HIPK2和药理学抑制剂。提出的研究将建立铁的分子机制 SH3RF1和HIPK2在肺成纤维细胞激活中的肺成纤维细胞和新作用中的积累。