tRNA-derived non-coding RNAs in ASM function and in asthma

tRNA 衍生的非编码 RNA 在 ASM 功能和哮喘中的作用

基本信息

批准号：
10643968
负责人：
Deepak A Deshpande
金额：
$ 58.81万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10643968
关键词：
Adhesions Allergic Allergic inflammation Anticodon Asthma Biogenesis Biological Markers Biological Models Bronchoconstriction Cell Proliferation Cell physiology Cells Cellular biology Data Development Enzymes Epithelial Cells Etiology Fibroblasts Focal Adhesions Gene Expression Gene Expression Regulation Goals Gonadal Steroid Hormones Growth Factor Human Inflammation Inflammation Mediators Inflammatory Infiltrate Inhalation Knowledge Ligation Link Lung Malignant Neoplasms Mediating Methodology Methods Molecular Morphology Mucous body substance Mus Muscle function Neurodegenerative Disorders Pathogenesis Pathology Pathway interactions Periodicity Phenotype Phosphorylation Plasma Play Process Production Proliferating Protein Tyrosine Kinase Proteins Pyroglyphidae RNA RNA analysis Regulation Regulator Genes Research Ribonucleases Role Sampling Signal Pathway Smooth Muscle Myocytes Stress Transcriptional Regulation Transfer RNA Translational Regulation Untranslated RNA Up-Regulation airway epithelium airway remodeling angiogenin asthmatic asthmatic patient biological adaptation to stress cell growth regulation cell motility chemokine chronic inflammatory disease chronic inflammatory lung disease cytokine experience individual patient inorganic phosphate insight migration novel novel therapeutics overexpression posttranscriptional prevent respiratory smooth muscle transcriptome transcriptome sequencing transcriptomic profiling

项目摘要

Project Summary and Abstract: The goal of our proposed studies is to elucidate the expression profiles and molecular functions of short non-coding RNAs (ncRNAs) in the pathogenesis of asthma. Asthma is a chronic inflammatory disease characterized by inflammation, mucus production, airway re-modeling, and hyper- responsiveness. In asthma, allergic inflammatory mediators, such as cytokines, act on resident airway cells [airway smooth muscle (ASM) cells, epithelial (AE) cells and fibroblasts (LF)] causing their structural and functional changes. However, knowledge gaps remain in our understanding of the mechanisms by which inflammatory mediators modulate cellular phenotypes. Although transcriptional regulation of gene expression in resident airway cells has been extensively studied, regulatory mechanisms at post-transcriptional steps remain elusive. In this context, short ncRNAs have evolved as one of the key post-transcriptional regulators of gene expression. Previous transcriptome profiling for short ncRNA analyses relied mainly on standard RNA-seq methods which fail to detect many RNA species. Cyclic phosphate-containing RNAs (cP-RNAs), that harbor a cyclic phosphate (cP) at their 3′-end, are one such RNA species not captured by RNA-seq, because cP prevents 3′-adapter ligation. Their absence in RNA-seq data makes cP-RNAs an invisible, hidden component of transcriptomes. Importantly, cP-RNAs are expressed as functional molecules. For example, angiogenin- generated 5′-tRNA halves, containing a cP and thus belonging to cP-RNAs, have functional significance in stress response, translational regulation, and cell proliferation, and are associated with neurodegenerative diseases and cancers. We propose that 5′-tRNA halves and other cP-RNAs play important roles in asthma pathobiology. In preliminary studies, we found that mouse lung expresses specific 5′-tRNA halves and cP-RNAs whose levels are upregulated during allergic inflammation caused by inhaled challenge of house dust mite (HDM). Furthermore, in human ASM cells, 5′-tRNA halves function to regulate cellular focal adhesion, migration, and proliferation. These results allowed us to hypothesize that inflammatory mediators upregulate the levels of 5′- tRNA halves/cP-RNAs, which contributes to airway cellular phonotypic changes in the molecular pathogenesis of asthma. By using our developed cP-RNA-seq, we propose to fully elucidate the regulation of the expression of 5′-tRNA halves/cP-RNAs mediated by asthmatic conditions in human lung and plasma samples and in human ASM cells, AE cells, and LFs (Aim 1). We further propose to assess the functional effects of 5′-tRNA halves/cP- RNAs on cellular focal adhesion, migration, proliferation, and morphology of ASM cells (Aim 2) and to investigate the molecular mechanisms underlying the functional effects of those RNAs (Aim 3). The proposed studies will reveal a novel ncRNA-engaged pathway in asthma pathogenesis and will support the exploration of biomarkers in asthma.

项目摘要和摘要：我们提出的研究的目的是阐明表达曲线和哮喘发病机理中短非编码RNA（NCRNA）的分子功能。哮喘是慢性炎症性疾病的特征是炎症，粘液产生，气道重新建模和超级疾病响应能力。在哮喘中，过敏性炎症介质（例如细胞因子）作用于居民气道细胞 [气道平滑肌（ASM）细胞，上皮（AE）细胞和成纤维细胞（LF）]导致其结构和功能变化。但是，知识差距仍然存在于我们对所在机制的理解中炎症介质调节细胞表型。尽管基因表达的转录调节居民气道细胞已广泛研究，在转录后步骤处的调节机制仍然存在难以捉摸。在这种情况下，简短的NCRNA已演变为基因转录后调节因子之一表达。用于短NCRNA分析的先前转录组分析主要依赖于标准RNA-seq 无法检测许多RNA物种的方法。含有循环磷酸RNA（CP-RNA），含有A 循环磷酸盐（CP）在其3'-End处是RNA-Seq未捕获的RNA，因为CP可防止 3'-适当的连接。它们在RNA-seq数据中的缺失使CP-RNA成为看不见的，隐藏的组成部分转录组。重要的是，CP-RNA表示为功能分子。例如，血管生成素 - 产生的5'-tRNA一半包含CP并因此属于CP-RNA，在应力中具有功能意义反应，翻译调节和细胞增殖，并与神经退行性疾病有关和癌症。我们建议5'-tRNA和其他CP-RNA在哮喘病理生物学中起重要作用。在初步研究中，我们发现小鼠肺表达了特定的5'-tRNA一半和CP-RNA的水平在因房屋尘螨（HDM）吸入挑战引起的过敏性炎症期间，被上调。此外，在人ASM细胞中，5'-tRNA的一半起作用，可调节细胞粘着粘合剂，迁移和增殖。这些结果使我们能够假设炎症介体上调了5'-的水平 tRNA半/CP-RNA，有助于分子发病机理的气道细胞音型变化哮喘。通过使用我们开发的CP-RNA-Seq，我们提议完全阐明表达的调节人类肺和血浆样品中哮喘条件介导的5'-tRNA半/CP-RNA以及人类 ASM细胞，AE细胞和LFS（AIM 1）。我们进一步建议评估5'-tRNA一半/cp-的功能效应 ASM细胞的细胞局灶性粘附，迁移，增殖和形态的RNA（AIM 2）并研究这些RNA功能效应的基础的分子机制（AIM 3）。拟议的研究将在哮喘发病机理中揭示了一种新型NCRNA参与的途径，并将支持生物标志物的探索在哮喘中。