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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

项目概要/摘要特发性肺纤维化 (IPF) 是一种与年龄相关的致命疾病，其特征为慢性、进行性和不可逆的纤维化。细胞衰老与 IPF 的发病机制广泛相关。然而，促进 IPF 衰老的机制，特别是在转录后水平，人们知之甚少。这当前的提案将通过阐明替代多聚腺苷酸化 (APA) 在衰老成纤维细胞及其对肺纤维化的贡献。 Nudix Hydrolase 21（Nudt21，也称为 CFIm25）是一种 RNA 结合蛋白，在 APA。我们最近发现 NDUT21 在衰老和纤维化的肺部以及衰老的肺部中表达下调成纤维细胞。正常肺成纤维细胞中 Nudt21 敲低诱导 STAT3 磷酸化和许多衰老相关的分泌表型（SASP）因素。重要的是，成纤维细胞 Nudt21 耗竭加剧博来霉素诱导的小鼠肺纤维化，而 NUDT21 拯救则导致纤维化减弱和 SASP 发布。这些发现强调 NUDT21 下调是一种新的可改变的衰老因素相关的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通过 APA 促进衰老过程中的肺纤维化 STAT3 信号传导和 SASP 诱导的调节。我们将在以下具体目标中检验这一假设：1）。评估 NUDT21 作为年龄相关性肺纤维化重要介质的作用； 2）确定衰老过程中 NUDT21 耗尽的上游机制； 3）定义针对NUDT21下游的作用肺纤维化的组成部分； 4) 人类 miRNA/NUDT21/STAT3 轴的分子研究 IPF 肺部。这些拟议的研究意义重大，因为它们将阐明一种新的机制，以增强 SASP 蛋白在衰老成纤维细胞中的表达可能有助于 IPF 的发病机制。创新包括证明 miRNA/NUDT21/STAT3/SASP 轴是衰老与衰老之间的新联系。 IPF 发病机制以及新试剂和方法，包括 Nudt21 敲除小鼠、原发性肺和来自老年/IPF 受试者的成纤维细胞，以及动物模型中的创新肺功能研究。最终结果这些研究将促进针对这种破坏性的与年龄相关的新治疗策略的开发疾病。