The role of small RNA derived tRNAs in gene regulation: Mechanism and Therapeutic Applications

小RNA衍生的tRNA在基因调控中的作用：机制和治疗应用

基本信息

批准号：
9365781
负责人：
Mark A Kay
金额：
$ 52.78万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2021-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9365781
关键词：
Affect Antisense Oligonucleotides Apoptosis Biogenesis Cell Death Cell Proliferation Cell Survival Cells Code Complement Data Defect Dose Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Genome Goals High-Throughput Nucleotide Sequencing Human Implant Leucine Literature Liver Liver neoplasms Malignant Neoplasms Mammalian Cell Mammals Measures Mediating Medical Messenger RNA Modeling Modernization Molecular Mus Oligonucleotides Organism Patients Phenotype Play Population Porifera Primary carcinoma of the liver cells Process Proteins RNA RNA, Ribosomal, 18S Regulation Regulator Genes Regulatory Pathway Ribosomal Proteins Ribosomes Role Safety Signal Transduction Small RNA Structure Testing Therapeutic Tissues Transfer RNA Translations Untranslated RNA Variant Xenograft procedure adeno-associated viral vector base cell growth clinical efficacy dark matter differential expression falls gene therapy human disease in vivo individual patient knock-down mammalian genome mouse model new therapeutic target novel pre-clinical response therapeutic target transgene expression tumor tumor growth

项目摘要

ABSTRACT One of the most important concepts in modern genetics is the realization that over 90% of the mammalian genome is transcribed into RNA. These non-protein coding RNAs have been described as the dark matter of the genome as the function of most of these RNAs has yet to be determined. Just over 5 years ago, we stumbled onto a novel set of tRNA derived small RNAs (tsRNAs). These tRNA derived fragments are also commonly called trfRNAs, and were originally thought to represent degradation products. However, it is now clear there are hundreds or perhaps thousands of different RNA fragments generated in mammalian cells and there is increasing evidence that these are functionally important. As a group, they have been proposed to regulate genes at many different levels. We plan to study the 3’tsRNAs (derived from the 3’ end of the mature tRNA) currently the least well studied of the various types of fragments, and for which no definitive function has yet to be described. We have established that one specific RNA, the 22nt CAG-Leucine 3’tsRNA when down regulated by the addition of antisense oligonucleotides in rapidly dividing cells inhibits ribosome biogenesis by first limiting the translation of at least one ribosomal protein mRNA. This results in rapid cellular apoptosis. In contrast, the addition of a 3’tsRNA mimic increases cellular proliferation and can complement the ribosome biogenesis defects in cells. These data bring to light a previously unknown post- transcriptional gene regulatory pathway. We plan to further explore the mechanism of how this specific 3’tsRNA-mediates translational gene regulation as well as use new high-throughput sequencing approaches to identify and then test other 3’tsRNAs that may participate in similar types of gene regulation. Finally, we propose to use gene therapy and oligonucleotide antisense delivery to pursue the therapeutic potential of manipulating 3’tsRNAs in treating medical conditions that affect the liver. We believe unraveling the molecular mechanism of this regulatory circuit is important both to further understand how genes are regulated and will provide novel therapeutic targets.

抽象的现代遗传学中最重要的概念之一是认识到超过 90% 的哺乳动物基因组被转录成RNA，这些非蛋白质编码RNA被描述为暗物质。就在 5 年前，我们还没有确定大多数这些 RNA 的基因组功能。偶然发现了一组新颖的 tRNA 衍生小 RNA (tsRNA)，这些 tRNA 衍生片段也是如此。通常称为 trfRNA，最初被认为代表降解产物，但现在它被认为是降解产物。显然，哺乳动物细胞中产生了数百甚至数千个不同的 RNA 片段越来越多的证据表明，它们作为一个整体具有重要的功能。我们计划研究 3'tsRNA（源自 3' 端）。成熟的 tRNA）目前是各种类型片段中研究最少的，并且没有明确的研究我们已经确定了一种特定的 RNA，即 22nt CAG-亮氨酸。当通过在快速分裂的细胞中添加反义寡核苷酸来下调 3'tsRNA 时，会抑制通过首先限制至少一种核糖体蛋白 mRNA 的翻译来实现核糖体生物发生。相比之下，添加 3'tsRNA 模拟物会增加细胞增殖，并且可以促进细胞凋亡。这些数据补充了细胞中核糖体生物发生的缺陷。我们计划进一步探讨这一具体的转录基因调控途径。 3’tsRNA 介导翻译基因调控以及使用新的高通量测序方法识别并测试可能参与类似类型基因调控的其他 3'tsRNA。提议使用基因疗法和寡核苷酸反义递送来追求治疗潜力我们相信操纵 3'tsRNA 来治疗影响肝脏的疾病。这种调节回路的分子机制对于进一步了解基因如何发挥作用非常重要受到监管并将提供新的治疗靶点。