The Role of RNA Binding Motif 5 in Traumatic Brain Injury

RNA 结合基序 5 在创伤性脑损伤中的作用

基本信息

批准号：
9494899
负责人：
TRAVIS C JACKSON
金额：
$ 34.23万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-01 至 2019-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9494899
关键词：
Apoptosis Autophagocytosis Axon Binding Brain Cardiovascular Diseases Caspase Cause of Death Cell Death Cells Cerebral Ischemia Cessation of life Cognitive Data Disease Down-Regulation Enterobacteria phage P1 Cre recombinase Etiology Event Exons Foundations Funding Future Gene Deletion Gene Expression Genes Hippocampus (Brain)Histologic Human Impairment In Vitro Injury Knock-out Knockout Mice Knowledge Learning Location Malignant Neoplasms Measures Mechanics Mediating Medical Memory Messenger RNA Modeling Molecular Monomeric GTP-Binding Proteins Motor Multiple Trauma Mus Nervous System Trauma Neurologic Neuronal Injury Neurons Normal Cell Nuclear Organ Outcome Pathologic Play Predisposition Proteins Publishing RBM5 gene RNA Binding RNA Splicing RNA analysis RNA-Binding Proteins Rattus Reporting Research Research Activity Resource Sharing Resources Role Signal Transduction Spinal Cord Spinal cord injury Staurosporine Stretching Subfamily lentivirinae Testing Time Tissues Toxic effect Trauma Traumatic Brain Injury United States National Institutes of Health Up-Regulation Vertebral column Work behavioral outcome cancer cell cell growth central nervous system injury clinically relevant cognitive function controlled cortical impact data resource disability drug development experimental study gene function improved in vitro testing in vivo in vivo evaluation innovation knock-down mutant nervous system disorder neurological recovery neuron loss neuropathology neuroprotection neurotoxic neurotoxicity next generation sequencing novel novel therapeutic intervention novel therapeutics overexpression prevent repaired small molecule tool transcriptome treatment strategy

项目摘要

ABSTRACT Traumatic brain injury (TBI) is a leading cause of death and disability in the young. Treatments are desperately needed to prevent cell death, repair neuronal connectivity, and improve cognitive outcomes after a TBI. This project will test if gene deletion of RNA binding motif 5 (RBM5) in mice improves molecular and histological readouts and behavioral outcomes after a controlled cortical impact (CCI) injury. RNA binding proteins (RBPs) regulate all aspects of mRNA (e.g. RNA splicing, gene expression, stability, and cellular localization). In recent years a number of RBPs have been associated with the etiology of disorders ranging from cancer, to cardiovascular and neurological disease. RBM5 is a pro-death RBP. The mechanisms by which RBM5 induces cell death involves upregulation of pro-death genes and also via the modulation of mRNA splicing ,which gives rise to toxic proteins with increased potency. Forced overexpression of RBM5 in cancer cells causes apoptosis, autophagy, and impedes cell growth. Very little is known about the function(s) of RBM5 in non-cancerous (i.e. normal) cells and tissues. Germane to the CNS, we showed that RBM5 levels increase after a TBI in mice. Independent of our work, RBM5 levels also reportedly increase after a traumatic spinal cord injury. It remains to be elucidated if RBM5 activates cell death in primary CNS cells (as it does in cancer). Our new preliminary data show that RBM5 overexpression in primary rat cortical neurons increases their susceptibility to a subsequent mechanical stretch-injury. This new finding is in line with our hypothesis that (1) RBM5 inhibition is a promising new therapeutic strategy for the treatment of CNS injury, and (2) suggests that RBM5 dysregulation may mediate pathological changes in gene expression which has been observed to occur after a TBI. In this project we will definitely test if RBM5 inhibition is neuroprotective in vivo. Novel conditional RBM5 KO mice will be subjected to a CCI induced injury. Neuropathology in WT versus KOs will be examined 1-21d post-injury. Also, learning and memory function will be examined 14-21d post-injury. Additionally we will analyze changes in global gene expression/splicing in primary cortical neurons after a mechanical stretch- injury, and also in which RBM5 levels are manipulated (i.e. by lentivirus mediated knockdown vs. overexpression). This study represents the most in-depth, largest, and only analysis of RBM5 gene function in brain which has been done to date.

抽象的创伤性脑损伤（TBI）是年轻人死亡和残疾的主要原因。治疗方法有迫切需要防止细胞死亡、修复神经元连接并改善认知结果创伤性脑损伤。该项目将测试小鼠中 RNA 结合基序 5 (RBM5) 的基因删除是否会改善分子和受控皮质撞击（CCI）损伤后的组织学读数和行为结果。 RNA 结合蛋白 (RBP) 调节 mRNA 的各个方面（例如 RNA 剪接、基因表达、稳定性和细胞定位）。近年来，许多 RBP 与以下疾病的病因有关：疾病范围从癌症到心血管和神经系统疾病。 RBM5 是一种支持死亡的 RBP。这 RBM5 诱导细胞死亡的机制涉及促死亡基因的上调，并且还通过 mRNA 剪接的调节，产生效力增强的有毒蛋白质。强迫过度表达癌细胞中的 RBM5 会导致细胞凋亡、自噬并阻碍细胞生长。人们对此知之甚少 RBM5 在非癌性（即正常）细胞和组织中的功能。 Germane 到 CNS，我们证明了小鼠 TBI 后 RBM5 水平升高。据报道，与我们的工作无关，RBM5 水平在外伤性脊髓损伤。 RBM5 是否激活原代 CNS 细胞的细胞死亡仍有待阐明（如癌症中确实如此）。我们的新初步数据表明 RBM5 在原代大鼠皮质神经元中过度表达增加了他们对随后的机械性拉伸损伤的敏感性。这一新发现符合我们的假设 (1) RBM5 抑制是一种有前途的治疗中枢神经系统损伤的新治疗策略， (2) 表明 RBM5 失调可能介导基因表达的病理变化，观察到发生在 TBI 后。在这个项目中，我们肯定会测试 RBM5 抑制是否具有体内神经保护作用。小说有条件 RBM5 KO 小鼠将遭受 CCI 诱导的损伤。将检查 WT 与 KO 中的神经病理学受伤后 1-21 天。此外，将在受伤后 14-21 天检查学习和记忆功能。另外我们将分析机械拉伸后原代皮质神经元整体基因表达/剪接的变化损伤，以及 RBM5 水平被操纵（即通过慢病毒介导的敲低与通过慢病毒介导的敲低）过度表达）。这项研究代表了对 RBM5 基因功能最深入、最大且唯一的分析。迄今为止已完成的大脑。