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

• 批准号: 10016850
• 负责人: TRAVIS C JACKSON
• 金额: $ 32.7万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10016850
• 关键词: 3-Dimensional; 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; Structure; Subfamily lentivirinae; TBI treatment; 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

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.

摘要 创伤性脑损伤（TBI）是年轻人死亡和残疾的主要原因。治疗是 迫切需要防止细胞死亡，修复神经元连接，并改善认知结果后， 创伤性脑损伤该项目将测试小鼠中RNA结合基序5（RBM 5）的基因缺失是否会改善分子和生物学特性。 组织学读数和受控皮质撞击（CCI）损伤后的行为结果。 RNA结合蛋白（RBP）调节mRNA的所有方面（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： 从癌症到心血管和神经系统疾病。RBM 5是一种亲死亡RBP。的 RBM 5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，其产生具有增加的效力的毒性蛋白。强迫过表达 RBM 5在癌细胞中的表达导致细胞凋亡、自噬并阻碍细胞生长。关于这一点，我们知之甚少。 RBM 5在非癌（即正常）细胞和组织中的功能。Germane到CNS，我们表明， 小鼠TBI后RBM 5水平增加。独立于我们的工作，据报道，RBM 5水平在 脊髓损伤RBM 5是否激活原代CNS细胞中的细胞死亡（如 在癌症中是这样的）。我们的新的初步数据表明，RBM 5在原代大鼠皮层神经元中的过表达 增加了他们对随后的机械拉伸损伤的易感性。这一新发现与我们的 假设（1）RBM 5抑制是治疗CNS损伤的有前景的新治疗策略， 和（2）表明RBM 5失调可能介导基因表达的病理变化， 在TBI后观察到。 在这个项目中，我们将明确测试RBM 5抑制是否在体内具有神经保护作用。新颖条件 RBM 5 KO小鼠将经受CCI诱导的损伤。将检查WT与科斯的神经病理学 伤后1- 21 d。另外，将在损伤后14- 21天检查学习和记忆功能。此外，我们将 分析机械拉伸后原代皮层神经元中整体基因表达/剪接的变化- 损伤，并且其中RBM 5水平被操纵（即，通过慢病毒介导的敲低vs. 过表达）。这项研究是迄今为止对RBM 5基因功能的最深入、最大和唯一的分析。 迄今为止已经完成的大脑。