Tiny RNAs as new potential biomarkers for gammaherpesvirus-driven neurological and central nervous system diseases

微小RNA作为伽马疱疹病毒驱动的神经和中枢神经系统疾病的新潜在生物标志物

• 批准号: 10727761
• 负责人: Kotaro Nakanishi
• 金额: $ 23.22万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-10 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10727761
• 关键词: Affinity; Antibodies; Biological Assay; Biological Markers; Body Fluids; Brain; Brain Injuries; Burkitt Lymphoma; Cell Line; Cells; Central Nervous System; Central Nervous System Diseases; Child; Data Set; Development; Diagnosis; Disease; Epstein-Barr Virus Infections; Foundations; Functional disorder; Gel; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Gene Silencing; Generations; Genes; Goals; Herpesviridae Infections; Hour; Human; Human Herpesvirus 4; Immunocompromised Host; Immunoprecipitation; Individual; Infection; Interferons; Label; Literature; Luciferases; Lymphocyte; Lymphoma cell; Lytic; Messenger RNA; MicroRNAs; Monitor; Mutation; Neurodevelopmental Disorder; Neurologic; Neurons; Nucleotides; Outcome; Paper; Pathogenesis; Patients; Pattern; Phosphodiesterase I; Plasma; Point Mutation; Proteins; PubMed; Publishing; RNA; RNA Interference; RNA-Induced Silencing Complex; Radiolabeled; Reporter; Reporting; Sepharose; Serum; Solid; Structure; System; Three Prime Repair Exonuclease 1; Transfection; Translational Repression; Virus Diseases; Virus Latency; Virus Replication; Work; anti-IgG; frontier; gammaherpesvirus; mRNA Translation; microRNA biomarkers; mutant; nervous system disorder; potential biomarker; reactivation from latency; transcriptome sequencing; vector

PROJECT SUMMARY Epstein-Barr virus (EBV) is known to damage the brain and central nervous system directly and indirectly through infected lymphocytes. These viruses are latent in healthy conditions but reactivate in immunocompromised individuals with severe consequences. Meanwhile, increasingly diverse literature has established that microRNAs (miRNAs) are extensively engaged in typical brain development, function, and dysfunction. In humans, 20~23- nucleotide (nt) miRNAs are loaded into four Argonaute (AGO) proteins, forming RNA-induced silencing complexes (RISCs) to repress the translation of mRNAs complementary to their miRNAs. Thus, proper gene regulation by miRNAs is indispensable for generating the neurological system, and their atypical expression patterns reflect the pathogenesis of neuronal diseases. Since miRNAs are known to circulate in serum, plasma, and other bodily fluids, they are used as biomarkers for diagnosing many diseases. In this context, we recently discovered that AGO- associated miRNAs are trimmed to 14-nt or shorter tiny RNAs (tyRNAs) by three 3′→5′ exonucleases: interferon- stimulated gene 20 kDa (ISG20), three prime repair exonuclease 1, and enhanced RNAi 1. ISG20 is highly expressed during EBV replication. Notably, our dual-luciferase reporter assay demonstrated that 14-nt tyRNAs no longer retain gene-silencing activity. These results suggest that viral infection globally converts miRNAs to tyRNAs, thereby making the gene expression drastically different. According to PubMed, the study of miRNAs has steadily increased, and more than 200,000 papers have already been published. In contrast, there have only been about 10 papers discussing tyRNAs, including our recent work, which demonstrates that little is known about tyRNAs. With the following two specific aims, we will determine the tyRNAs in the four different stages of EBV-infected cells: 1) no infection, 2) de novo infection, 3) latency, and 4) lytic reactivation (Aim 1). In addition, we will validate the hypothesis that the herpesvirus infection enhances tyRNA generation in neurodevelopmental disorder patients whose AGO has specific single-point mutations (Aim 2). The successful outcome of the proposed study will provide the first comprehensive data sets of tyRNAs as new potential biomarkers. The long-term goal of this study is to provide a foundation to establish a new frontier in understanding how herpesvirus infection generates tyRNAs and thus dysregulates gene expression. Obtaining the comprehensive profile of tyRNAs in the proposed project is an essential first step to proceeding towards a deeper study. The determined tyRNAs are expected to serve as biomarkers and make previously identified miRNA biomarkers more informative.

项目摘要 已知Epstein-Barr病毒（EBV）直接和间接地损害了大脑和中枢神经系统 感染的淋巴细胞。这些病毒在健康条件下潜在潜伏期中，但在免疫功能下重新激活 有严重后果的人。同时，越来越多样化的文献已经确定了microRNA （miRNA）广泛参与典型的大脑发育，功能和功能障碍。在人类中，20〜23- 将核苷酸（NT）miRNA加载到四个Argona（AGO）蛋白中，形成RNA诱导的沉默复合物 （riscs）反映mRNA互补为miRNA的翻译。那是通过 miRNA对于产生神经系统是必不可少的，它们的非典型表达模式反映了 神经元疾病的发病机理。由于已知miRNA在血清，血浆和其他体液中循环，所以 它们被用作诊断许多疾病的生物标志物。在这种情况下，我们最近发现了Ago- 相关的miRNA通过三个3'→5'外核酸酶修剪为14-nt或较短的小RNA（Tyrnas）：干扰素 - 受刺激的基因20 kDa（ISG20），三个素修复外切核酸酶1和增强的RNAi1。SG20高度表达 在EBV复制期间。值得注意的是，我们的双脂肪酸酶报道器测定法证明了14-nt Tyrnas不再保留 基因分解活性。这些结果表明，全球病毒感染将miRNA转化为泰尔纳（Tyrnas），从而使 基因表达截然不同。根据PubMed的说法，对miRNA的研究已悄悄增加，并且 已经发表了200,000多篇论文。相反，只有大约10篇论文 讨论Tyrnas，包括我们最近的作品，这表明对Tyrnas知之甚少。与 以下两个具体目标，我们将在EBV感染的细胞的四个不同阶段确定Tyrnas：1）否 感染，2）从头感染，3）潜伏期和4）裂解重新激活（AIM 1）。另外，我们将验证假设 疱疹病毒感染可增强tyrna的产生 特定的单点突变（AIM 2）。拟议研究的成功结果将提供第一个 Tyrnas作为新的潜在生物标志物的全面数据集。这项研究的长期目标是提供 基础建立一个新的边界，以了解疱疹病毒感染如何产生泰尔纳人，从而 失调基因表达。在拟议项目中获得Tyrnas的全面概况是必不可少的 进行更深入研究的第一步。确定的泰尔纳人有望用作生物标志物，并且 使先前确定的miRNA生物标志物更有用。