Controlling toxic RNA with rapamycin

用雷帕霉素控制有毒 RNA

基本信息

批准号：
8676619
负责人：
Jeremy Robert Sanford
金额：
$ 19.13万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA CRUZ
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8676619
关键词：
Address Aging Alternative Splicing Antisense Oligonucleotides Area Autophagocytosis Binding Biochemical Bioinformatics Biological Biological Assay Biology of Aging Boxing California Cell Aging Cell Nucleus Commit Complex Computational Biology Data Diabetes Mellitus Diploid Cells Disease Environment Equilibrium Event Functional disorder Future Gene Expression Gene Expression Regulation Gene Family Genes Genetic Translation Genomics Goals Human Laboratories Link Longevity Macrolides Malignant Neoplasms Mediating Messenger RNA Methods MicroRNAs Molecular Molecular Biology Nonsense Codon Nonsense-Mediated Decay Nuclear Oncogenic Oranges Pathway interactions Phosphorylation Play Post-Transcriptional Regulation Postdoctoral Fellow Process Protein Isoforms Proteins Publications Quality Control RNA RNA Splicing Regulation Regulator Genes Relative (related person)Research Research Project Grants Role Signal Transduction Sirolimus Therapeutic Transcript Translations Universities Vision base innovation interest longevity gene mRNA Precursor mRNA Stability mRNA Surveillance mRNA capping mTOR protein novel prevent public health relevance research study transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): The goal of this proposal is to explore novel connections between the rapamycin/mTOR pathway, non-sense mediated decay (NMD) and post- transcriptional control of gene expression. We propose that by enhancing NMD, rapamycin contributes to degradation of aberrant mRNAs containing premature termination codons (PTCs). In Aim 1 we propose four different, non-exclusive hypotheses by which rapamycin may enhance NMD. The goal of this exploratory R21 proposal is to invalidate at least one of these hypotheses, enabling us to focus on those mechanisms most relevant to aging and post- transcriptional gene regulation. In hypothesis 1, we will determine if the mTORC1 pathway regulates NMD. In hypothesis 2 we propose that rapamycin/mTOR regulates the expression, phosphorylation and/or localization of the NMD machinery. Hypothesis 3 proposes that rapamycin may influence the equilibrium between the pioneer and steady rounds of translation. Finally, in hypothesis 4 we propose that rapamycin/mTOR may modulate NMD efficiency by directly regulating pre-mRNA splicing decisions in the nucleus. Collectively, aim 1 will reveal a novel mechanism by which rapamycin/mTOR modulates NMD and thereby the decay of toxic mRNAs. In Aim 2 we will employ high throughput RNA-Seq to determine how rapamycin/mTOR coordinates the expression of gene regulatory networks associated with cellular aging. Our preliminary results shown that rapamycin/mTOR coordinates AS and translation of specific isoforms. Employing an innovative sequencing method, developed in our lab, and bioinformatics we will identify gene families regulated at the level of splicing, mRNA stability and translation b rapamycin/mTOR. Aim 2 will reveal novel candidates for future use in RNA-based therapeutics employing antisense-oligonucleotides to modulate specific gene isoforms processing and promote splicing and translation of anti-oncogenic and pro-longevity genes.

描述（由申请人提供）：该建议的目的是探索雷帕霉素/mTOR途径，非义介导衰减（NMD）和基因表达的转录后控制之间的新连接。我们建议，通过增强NMD，雷帕霉素有助于含有过早终止密码子（PTC）的异常mRNA降解。在AIM 1中，我们提出了雷帕霉素可以增强NMD的四个不同的非排他性假设。该探索性R21提案的目的是使至少这些假设中的至少一个无效，从而使我们能够专注于与衰老和转录后基因调控最相关的这些机制。在假设1中，我们将确定MTORC1途径是否调节NMD。在假设2中，我们提出雷帕霉素/mTOR调节NMD机械的表达，磷酸化和/或定位。假设3提出，雷帕霉素可能会影响先锋和稳定的翻译圈之间的平衡。最后，在假设4中，我们提出，雷帕霉素/mTOR可能通过直接调节细胞核中的mRNA剪接决策来调节NMD效率。总体而言，AIM 1将揭示一种新型机制，雷帕霉素/mTOR调节NMD，从而使有毒mRNA衰减。在AIM 2中，我们将采用高吞吐量RNA-Seq来确定雷帕霉素/mTOR如何协调与细胞衰老相关的基因调节网络的表达。我们的初步结果表明，雷帕霉素/mTOR坐标AS和特定同工型的翻译。采用在我们的实验室中开发的创新测序方法和生物信息学，我们将确定在剪接，mRNA稳定性和翻译水平上调节的基因家族B雷帕霉素/mTOR。 AIM 2将揭示使用反义 - 寡核苷酸来调节特定基因同工型加工并促进抗康复和促longevity基因的剪接和翻译的新型候选者，以在基于RNA的治疗剂中使用。