Regulatory role of APA in pulmonary fibrosis during aging

APA在衰老过程中肺纤维化中的调节作用

• 批准号: 10674253
• 负责人: Zheng Chen
• 金额: $ 31.98万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10674253
• 关键词: 3' Untranslated Regions; Address; Age; Aging; Animal Model; Attenuated; Binding; Bleomycin; Bypass; Cell Aging; Chronic; Collagen; Data; Development; Disease; Distal; Down-Regulation; Effector Cell; Epithelial Cells; Fibroblasts; Fibrosis; Foundations; Genes; Genetic Transcription; Human; In Vitro; Interleukins; Investigation; Knockout Mice; Knowledge; Lead; Length; Link; Lung; Matrix Metalloproteinases; Mediating; Mediator of activation protein; Methodology; MicroRNAs; Modeling; Molecular; Mus; Myofibroblast; Names; Oligonucleotides; Pathogenesis; Patients; Phenotype; Phosphorylation; Play; Poly(A) Tail; Polyadenylation; Population; Proteins; Pulmonary Fibrosis; RNA; RNA-Binding Proteins; Reagent; Regulation; Role; STAT3 gene; Seminal; Signal Transduction; Site; Testing; Transcript; Translating; age related; aged; alveolar epithelium; base; defined contribution; experimental study; fibrotic lung; idiopathic pulmonary fibrosis; in vivo; innovation; knock-down; mouse model; novel; novel therapeutic intervention; nudix hydrolase; overexpression; prevent; response; senescence; single-cell RNA sequencing

PROJECT SUMMARY/ABSTRACT Idiopathic Pulmonary Fibrosis (IPF) is a lethal age-related disease characterized by chronic, progressive, and irreversible fibrosis. Cellular senescence has been widely implicated in the pathogenesis of IPF. However, the mechanism promoting senescence in IPF, especially at the post-transcriptional level, is poorly understood. The current proposal will address this knowledge gap by elucidating the role of alternative polyadenylation (APA) in senescent fibroblasts and its contribution to pulmonary fibrosis. Nudix Hydrolase 21 (Nudt21, also known as CFIm25) is an RNA binding protein playing an important role in APA. We recently found that NDUT21 is downregulated in aging and fibrotic lungs as well as in senescent fibroblasts. Nudt21 knockdown in normal lung fibroblasts induces STAT3 phosphorylation and the expression of many senescence-associated secretory phenotype (SASP) factors. Importantly, fibroblast Nudt21 depletion aggravated bleomycin-induced pulmonary fibrosis in mice, whereas NUDT21 rescue led to attenuated fibrosis and SASP release. These findings highlight NUDT21 downregulation as a novel modifiable factor for aging- related IPF and provide a strong foundation to study the role of NUDT21-mediated-APA in lung fibrosis during aging. Based on these extensive preliminary findings, we hypothesize that miRNA-mediated NUDT21 downregulation in senescent fibroblasts promotes pulmonary fibrosis during aging, through APA regulation of STAT3 signaling and SASP induction. We will test this hypothesis in following specific aims: 1). Evaluate the role of NUDT21 as an important mediator in age-associated lung fibrosis; 2) Determine the upstream mechanism for NUDT21 depletion during aging; 3) Define the role of targeting NUDT21 downstream components in pulmonary fibrosis; and 4) Molecular investigation of the miRNA/NUDT21/STAT3 axis in human IPF lungs. These proposed studies are significant because they will elucidate a novel mechanism for enhanced expression of SASP proteins in senescent fibroblasts that could contribute to the pathogenesis of IPF. Innovations include demonstrating the miRNA/NUDT21/STAT3/SASP axis as a novel link between aging and IPF pathogenesis, and new reagents and methodologies including Nudt21 knockout mice, primary lungs and fibroblasts from aged/IPF subjects, and innovative lung functional studies in animal models. Ultimately, results from these studies will facilitate development of novel therapeutic strategies against this devastating age-related disease.

项目摘要/摘要 特发性肺纤维化（IPF）是一种致命的年龄相关疾病，其特征是慢性，进行性和进行性疾病 不可逆的纤维化。细胞衰老已广泛与IPF的发病机理有关。但是， 促进IPF衰老的机制，尤其是在转录后水平上的机制知之甚少。这 当前的建议将通过阐明替代聚腺苷酸化（APA）在 衰老成纤维细胞及其对肺纤维化的贡献。 NUDIX水解酶21（NUDT21，也称为CFIM25）是RNA结合蛋白在 APA。最近，我们发现NDUT21在衰老和纤维化肺以及衰老中被下调 成纤维细胞。正常肺成纤维细胞中的NUDT21敲低诱导Stat3磷酸化和表达 许多与衰老相关的分泌表型（SASP）因素。重要的是，成纤维细胞NUDT21耗竭 博来霉素诱导的小鼠肺纤维化的加重，而NUDT21救援导致纤维化减弱 和SASP释放。这些发现凸显了NUDT21的下调是衰老的新型可改变因素。 相关的IPF并为研究NUDT21介导的APA在肺纤维化中的作用提供了坚实的基础 老化。基于这些广泛的初步发现，我们假设miRNA介导的NUDT21 衰老成纤维细胞的下调可在衰老期间促进肺纤维化，通过APA STAT3信号传导和SASP诱导的调节。我们将以以下特定目的检验该假设：1）。 评估NUDT21在与年龄相关的肺纤维化中的重要介体的作用； 2）确定 衰老期间NUDT21耗竭的上游机制； 3）定义靶向下游NUDT21的作用 肺纤维化中的成分； 4）人类中miRNA/NUDT21/STAT3轴的分子研究 IPF肺。这些提出的研究很重要，因为它们将阐明一种新的机制来增强 SASP蛋白在衰老成纤维细胞中的表达，可能有助于IPF的发病机理。 创新包括证明miRNA/nudt21/stat3/sasp轴作为衰老与 IPF发病机理以及包括NUDT21敲除小鼠，原发性肺和包括NUDT21敲除小鼠的新试剂和方法论 来自老年/IPF受试者的成纤维细胞以及动物模型中创新的肺功能研究。最终，结果 从这些研究中将有助于发展与这种毁灭性年龄有关的新型治疗策略 疾病。