The mRNA splicing factor RBM5: A new therapeutic target for TBI

mRNA剪接因子RBM5：TBI的新治疗靶点

基本信息

批准号：
8845278
负责人：
TRAVIS C JACKSON
金额：
$ 23.1万
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：
2014
资助国家：
美国
起止时间：
2014-07-01 至 2016-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8845278
关键词：
Acute Brain Injuries Anthraquinones Apoptotic Applications Grants Behavioral Brain Brain Diseases Brain Injuries CASP8 and FADD-like apoptosis regulating protein CASP8 gene Caspase Cause of Death Cell Death Cells Cellular Stress Cessation of life Clinical Trials Cognitive Data Drosophila pros protein Drug Targeting Enhancers Enzymes Excision Exclusion Exons Genes Health Hippocampus (Brain)Human Hydrogen Peroxide Impairment In Vitro Injury Intervention Introns Knowledge Learning Length Lesion Mechanics Mediating Memory Messenger RNA Modeling Mus Neurons Nuclear Nuclear Proteins Pharmaceutical Preparations Play Population Process Protein Isoforms Protein Splicing Proteins Publications RNA Binding RNA Recognition Motif RNA Splicing Rattus Recovery Reporting Role Site Spliceosome Assembly Pathway Spliceosomes Staurosporine Stretching Structure Sulfonic Acids TNF gene TNFSF10 gene TNFSF6 gene Testing Testis Therapeutic Toxic effect Transcript Traumatic Brain Injury Up-Regulation Variant Zinc Fingers cancer cell caspase-2 central nervous system injury controlled cortical impact disability in vitro testing in vivo inhibitor/antagonist injured intravenous administration mRNA Expression mRNA Precursor new therapeutic target novel overexpression pro-caspase-2 protein B protein expression response small molecule social

项目摘要

DESCRIPTION (provided by applicant): Reports estimate that over 90% of all human genes are subject to mRNA splicing. RNA splicing occurs in highly specialized structures termed nuclear spliceosomes. Spliceosomes consist of a few key proteins that catalyze the excision of target introns/exons, plus 100's of co-factors that regulate the activity/efficiency of these enzymes. Spliceosome co-factors direct many facets of splicing dynamics including distribution of different protein variants throughout the body. Brain injury and disease disturb splicing. Pathological changes in spliceosome mechanics alter splice variant expression of different survival and death proteins. An upregulation of maladaptive protein variants may exacerbate neuronal death and impair CNS recovery. RNA Binding Motif 5 (RBM5) is a splicing co-factor. It is highly expressed in the CNS and testis but its function in the healthy or injured brain is unknown. RBM5 briefly localizes to the spliceosome, very early during spliceosome assembly around pre-mRNA targets. A zinc finger domain (RanBP2-Type), and several other RNA binding domains, coordinate its highly selective interaction with spliceable exons in pre-mRNA targets. In cancer cells, RBM5 regulates splice variant selection of caspase-2 (pro-death) and c-FLIP (pro-survival) genes. It promotes the exclusion of exon 9 from caspase-2, and exon 7 from c-FLIP mRNA. This induces caspase2L and c-FLIPL protein expression, respectively. Caspase2L is a pro-death splice variant. The c-FLIPL splice variant also has pro-apoptotic functions. In contrast, RBM5 inhibition causes exon 9/7 retention, respectively, and forces caspase-2s/c-FLIPs expression. Caspase-2s and c-FLIPs are both potent pro-survival variants. Thus RBM5 promotes cell death by upregulating the ratio of pro-death to pro-survival splice variants. RBM5 has not been studied in the brain. High expression in the CNS suggests that it may play a key role in splicing-mediated cell death mechanisms. To the best of our knowledge, no drug has ever been developed to specifically target pro-death splicing mechanisms in the brain - thus the therapeutic value of splicing directed therapies is completely unknown. Our preliminary data show that RBM5 is highly enriched in the hippocampal CA3 formation - a neuron population that is especially vulnerable to traumatic brain injury (TBI). Furthermore, we show that anthraquinone-2-sulfonic acid (AQ2S), the world's first small-molecule RBM5 inhibitor (that blocks the RanBP2-Type domain), decreases neuronal death after TBI in vitro and in vivo. This grant proposal seeks to test our hypothesis that RBM5 is a potent pro-death splicing factor in rat/human neurons (i.e. show that it upregulates caspase-2L and c-FLIPL splicing in these cells), and further confirm our preliminary data showing that AQ2S is the first viable drug to block pro-death splicing mechanisms in the brain.

描述（由申请人提供）：报告估计，超过 90% 的人类基因都受到 mRNA 剪接的影响。 RNA 剪接发生在称为核剪接体的高度专业化结构中。剪接体由一些催化靶内含子/外显子切除的关键蛋白质，以及调节这些酶的活性/效率的数百个辅助因子组成。剪接体辅助因子指导剪接动力学的许多方面，包括不同蛋白质变体在体内的分布。脑损伤和疾病会干扰剪接。剪接体力学的病理变化改变了不同存活和死亡蛋白的剪接变体表达。适应不良蛋白变异的上调可能会加剧神经元死亡并损害中枢神经系统的恢复。 RNA 结合基序 5 (RBM5) 是一种剪接辅助因子。它在中枢神经系统和睾丸中高度表达，但其在健康或受伤大脑中的功能尚不清楚。 RBM5 在剪接体围绕前 mRNA 靶标组装的早期阶段短暂定位于剪接体。锌指结构域（RanBP2 型）和其他几个 RNA 结合结构域协调其与前 mRNA 靶标中的可剪接外显子的高度选择性相互作用。在癌细胞中，RBM5 调节 caspase-2（促死亡）和 c-FLIP（促生存）基因的剪接变体选择。它促进从 caspase-2 中排除外显子 9，从 c-FLIP mRNA 中排除外显子 7。这分别诱导 caspase2L 和 c-FLIPL 蛋白表达。 Caspase2L 是一种促死亡剪接变体。 c-FLIPL 剪接变体还具有促凋亡功能。相反，RBM5 抑制分别导致外显子 9/7 保留，并强制 caspase-2s/c-FLIPs 表达。 Caspase-2 和 c-FLIP 都是有效的促生存变体。因此，RBM5通过上调促死亡与促存活剪接变体的比率来促进细胞死亡。 RBM5 尚未在大脑中进行研究。中枢神经系统中的高表达表明它可能在剪接介导的细胞死亡机制中发挥关键作用。据我们所知，还没有开发出专门针对大脑中促死亡剪接机制的药物，因此剪接定向疗法的治疗价值完全未知。我们的初步数据表明，RBM5 在海马 CA3 结构中高度富集，海马 CA3 是一个特别容易受到创伤性脑损伤 (TBI) 影响的神经元群。此外，我们还发现，世界上第一个小分子 RBM5 抑制剂（阻断 RanBP2 型结构域）蒽醌-2-磺酸 (AQ2S) 在体外和体内均可减少 TBI 后的神经元死亡。该拨款提案旨在检验我们的假设，即 RBM5 是大鼠/人类神经元中有效的促死亡剪接因子（即表明它上调这些细胞中的 caspase-2L 和 c-FLIPL 剪接），并进一步证实我们的初步数据表明AQ2S 是第一种阻断大脑促死亡剪接机制的可行药物。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

BrainPhys® increases neurofilament levels in CNS cultures, and facilitates investigation of axonal damage after a mechanical stretch-injury in vitro.

BrainPhys® 增加中枢神经系统培养物中的神经丝水平，并有助于体外机械拉伸损伤后轴突损伤的研究。

DOI：
发表时间：
2018-02
期刊：
影响因子：
5.3
作者：
Jackson, Travis C;Kotermanski, Shawn E;Jackson, Edwin K;Kochanek, Patrick M
通讯作者：
Kochanek, Patrick M

Cold stress protein RBM3 responds to temperature change in an ultra-sensitive manner in young neurons.

冷应激蛋白 RBM3 在年轻神经元中以超敏感的方式响应温度变化。

DOI：
发表时间：
2015-10-01
期刊：
影响因子：
3.3
作者：
Jackson, T C;Manole, M D;Kotermanski, S E;Jackson, E K;Clark, R S B;Kochanek, P M
通讯作者：
Kochanek, P M

The nuclear splicing factor RNA binding motif 5 promotes caspase activation in human neuronal cells, and increases after traumatic brain injury in mice.

核剪接因子 RNA 结合基序 5 促进人类神经元细胞中的 caspase 激活，并在小鼠脑外伤后增加。

DOI：
发表时间：
2015-03-31
期刊：
影响因子：
0
作者：
Jackson, Travis C;Du, Lina;Janesko;Vagni, Vincent A;Dezfulian, Cameron;Poloyac, Samuel M;Jackson, Edwin K;Clark, Robert S B;Kochanek, Patrick M
通讯作者：
Kochanek, Patrick M

It might be time to let cooler heads prevail after mild traumatic brain injury or concussion.

在轻度脑外伤或脑震荡之后，也许是时候让冷静的头脑占上风了。