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标是阐明表达谱和短非编码 RNA (ncRNA) 在哮喘发病机制中的分子功能。以炎症、粘液产生、气道重塑和过度呼吸为特征的炎症性疾病在哮喘中，过敏性炎症介质（例如细胞因子）作用于气道细胞。 [气道平滑肌 (ASM) 细胞、上皮 (AE) 细胞和成纤维细胞 (LF)] 导致其结构和然而，我们对功能变化的机制的理解仍然存在知识差距。炎症介质调节细胞表型，但基因表达的转录调节。驻留气道细胞已被广泛研究，转录后步骤的调节机制仍然存在在这种情况下，短 ncRNA 已进化为基因的关键转录后调节因子之一。以前用于短 ncRNA 分析的转录组分析主要依赖于标准 RNA-seq。无法检测许多含有环状磷酸盐的 RNA (cP-RNA) 的方法。 3' 端的环状磷酸盐 (cP) 是一种未被 RNA-seq 捕获的 RNA 种类，因为 cP 阻止 RNA-seq 数据中 3′-接头连接的缺失使得 cP-RNA 成为不可见的隐藏组件。重要的是，cP-RNA 表达为功能分子，例如血管生成素。生成的 5′-tRNA 半部含有 cP，因此属于 cP-RNA，在应激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相关我们认为 5'-tRNA 半体和其他 cP-RNA 在哮喘病理学中发挥着重要作用。在初步研究中，我们发现小鼠肺表达特定的 5′-tRNA 半体和 cP-RNA，其水平在吸入屋尘螨 (HDM) 引起的过敏性炎症期间上调。此外，在人 ASM 细胞中，5'-tRNA 的一半功能是调节细胞粘着斑、迁移和这些结果使我们能够对抗炎症介质上调 5'- 水平的问题。 tRNA half/cP-RNA，有助于分子发病机制中气道细胞表型的变化通过使用我们开发的 cP-RNA-seq，我们建议充分阐明表达的调节。人肺和血浆样本以及人体内哮喘状况介导的 5'-tRNA 半/cP-RNA ASM 细胞、AE 细胞和 LF（目标 1）我们进一步建议评估 5'-tRNA half/cP- 的功能效果。 RNA 对 ASM 细胞的细胞粘着斑、迁移、增殖和形态的影响（目标 2）并进行研究这些 RNA 功能作用的分子机制（目标 3）。揭示了哮喘发病机制中一条新的 ncRNA 参与途径，并将支持生物标志物的探索在哮喘中。