Novel approaches to identify regulators of Bak1 splicing

识别 Bak1 剪接调节因子的新方法

基本信息

批准号：
10040977
负责人：
Liang Chen
金额：
$ 45.29万
依托单位：
UNIVERSITY OF CALIFORNIA RIVERSIDE
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：
2020-08-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10040977
关键词：
Ablation Alternative Splicing Apoptosis Apoptotic Attenuated BAK1 gene Big Data Biochemical Brain CRISPR screen Cell Death Cell Line Cell Survival Cells Cessation of life Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Competence Computational Biology Coupled Cues Data Set Development Disease Exons Fluorescence Gene Expression Genetic Genetic Transcription Goals Homeostasis Investigation Knock-out Learning Life Longevity Mediating Memory Messenger RNA Methods MicroRNAs Mitochondrial Proteins Molecular Molecular Genetics Mus Natural regeneration Neurobiology Neurodegenerative Disorders Neurodevelopmental Disorder Neuronal Differentiation Neurons Nuclear Pore Complex Organism Pathologic Pathway interactions Physiological Positioning Attribute Predisposition Protein Isoforms Proteins Proteomics Public Health RNA RNA Splicing RNA-Binding Proteins Recording of previous events Regulation Regulatory Element Reporter Research Resistance Surveys Time Tissues Transcript Translations base brain tissue data mining knock-down mRNA Decay mortality nerve stem cell neural circuit neuron apoptosis neuron loss neuronal survival novel novel strategies postnatal pro-apoptotic protein programs protein expression relating to nervous system screening stable cell line tool transcriptome transcriptome sequencing transcriptomics

项目摘要

The objective of this application is to unbiasedly identify regulators essential for programming intrinsic neuronal resistance to apoptosis. Apoptosis is a ubiquitous regulated cell death pathway controlling cell turnover and tissue homeostasis in metazoans. A longstanding issue is how neurons suppress apoptosis in favor of longevity. Previous studies on neuronal regulation of apoptosis have focused on why and how extrinsic survival cues help establish and maintain neural circuits through the control of cell death. While these inspiring studies delineate how neurons compete for survival at the time of circuit formation, important questions remain unresolved, i.e., whether neuronal apoptosis sensitivity is genetically determined. In our preliminary studies, we have found that neuronal resistance to apoptosis is intrinsically programmed before circuit formation by depletion of pro-apoptotic mitochondrial protein BAK1. We further found that BAK1 expression is controlled at the RNA level through robust alternative splicing mechanisms. The critical question is: what factors program Bak1 splicing? Identification of these factors will have significant impacts and enable new investigations of neuronal cell death controls in various settings. The proposed study will generate new tools and develop robust cell-based methods to systematically identify regulators of Bak1 splicing. We will integrate experimental and computational approaches to accelerate discoveries that would otherwise be limited and less sensitive. Our team has demonstrated successful collaborations researching cell death, neuronal survival, and RNA molecular genetics in the brain, and expect the proposed research to be fruitful. By revealing novel regulatory mechanisms of apoptosis and associated genetic factors, our findings may inform strategies for enhancing cell survival and tackling neurodegenerative diseases. Completion of the proposed study will also provide proof-of-principle for our broadly applicable strategy to study any alternative exons.

该应用的目的是公正地识别对编程内在神经元必不可少的调节剂对凋亡的抗性。凋亡是一种无处不在的细胞死亡途径，控制细胞周转和后生动物的组织稳态。一个长期存在的问题是神经元如何抑制细胞凋亡而有利于长寿。先前关于凋亡神经元调节的研究集中于为什么以及外在生存如何通过控制细胞死亡，提示有助于建立和维持神经回路。而这些鼓舞人心的研究描述在电路形成时神经元如何争夺生存，重要的问题仍然存在未分辨，即神经元凋亡敏感性是否是遗传确定的。在我们的初步研究中，我们已经发现，神经元对凋亡的抗性是在电路形成之前本质上编程的促凋亡线粒体蛋白BAK1的耗竭。我们进一步发现Bak1表达在 RNA通过可靠的替代剪接机制的水平。关键问题是：哪些因素计划 bak1剪接？这些因素的识别将产生重大影响，并可以对神经元细胞死亡在各种环境中控制。拟议的研究将生成新工具并开发出强大的基于细胞的方法，可以系统地识别BAK1剪接的调节剂。我们将整合实验和加速发现的计算方法，否则将受到限制和敏感的敏感性。我们的团队已经证明了研究细胞死亡，神经元生存和RNA的成功合作大脑中的分子遗传学，并期望拟议的研究成果。通过揭示新的调节细胞凋亡的机制和相关的遗传因素，我们的发现可能会为增强细胞的策略提供信息生存和解决神经退行性疾病。拟议的研究的完成还将为我们的广泛适用策略提供原则上的研究，以研究任何替代外显子。