Antisense peptide nucleic acid targeting GluR3 delays disease onset and progression in the SOD1 G93A mouse model of familial ALS
Antisense peptide nucleic acid targeting GluR3 delays disease onset and progression in the SOD1 G93A mouse model of familial ALS
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DOI：
10.1002/jnr.20191
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发表时间：
2004-08-15
影响因子：
4.2
通讯作者：
Cheema, SS
中科院分区：
文献类型：
--
作者：
Rembach, A;Turner, BJ;Cheema, SS
谷氨酸兴奋性毒性是肌萎缩侧索硬化症（ALS）运动神经元变性的一个主要影响因素。兴奋性毒性由升高的细胞内钙离子（Ca 2+）水平引起，其进而募集细胞死亡信号传导途径。最近的证据表明，α-氨基-3-羟基-5-甲基-4-异恶唑丙酸（AMPA）受体亚单位（GluR）的化学计量是一个主导因素，导致过量的钙离子负载在神经退行性病变。特别是，Ca 2+渗透性谷氨酸受体亚基3（GluR 3）已涉及几种神经系统疾病，如双相情感障碍和癫痫。最近在我们的小组对铜锌超氧化物歧化酶（SOD 1）（G93 A）的转基因小鼠模型的家族性ALS（FALS）的蛋白质组学分析揭示了一个潜在的有害上调谷氨酸受体3在脊髓相比，在野生型同窝出生。基于这一发现，我们设计了一种针对GluR 3的12聚体反义肽核酸（PNA）。该序列显著降低了GluR 3蛋白水平，并保护神经母细胞瘤X脊髓（NSC-34）细胞免受AMPA受体特异性激动剂（S）-5-fluorowillardiine诱导的死亡。我们随后在出生后第50天开始每周三次用腹膜内注射反义PNA（2.5mg/kg）处理SOD 1G 93 A小鼠。与注射无义序列的小鼠相比，用反义序列处理的小鼠具有显著延长的存活。然而，蛋白质印迹分析没有显示腰椎脊髓的整个提取物中GluR 3蛋白水平显著降低。这些结果表明，干扰AMPA受体组装的GluR 3组分可能是控制运动神经元兴奋性毒性破坏的一种新策略，并可能为治疗人类ALS带来新的治疗机会。(C)2004 Wiley-Liss，Inc.
Glutamate excitotoxicity is strongly implicated as a major contributing factor in motor neuron degeneration in amyotrophic lateral sclerosis (ALS). Excitotoxicity results from elevated intracellular calcium ion (Ca2+) levels, which in turn recruit cell death signaling pathways. Recent evidence suggests that alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor subunit (GluR) stoichiometry is a dominant factor leading to excess Ca2+ loading in neurodegeneration. In particular, the Ca2+ permeable glutamate receptor subunit 3 (GluR3) has been implicated in several neurologic conditions such as bipolar disorder and epilepsy. Recent proteomic analysis within our group on the copper zinc superoxide dismutase (SOD1)(G93A) transgenic mouse model of familial ALS (FALS) reveals a potentially deleterious upregulation of GluR3 in spinal cord compared to that in wild-type littermates. Based on this finding we designed a 12mer antisense peptide nucleic acid (PNA) directed against GluR3. This sequence significantly reduced levels of GluR3 protein and protected neuroblastoma X spinal cord (NSC-34) cells against death induced by the AMPA receptor-specific agonist (S)-5-fluorowillardiine. We subsequently treated SOD 1 G93A mice thrice weekly with intraperitoneal injections of the antisense PNA (2.5 mg/kg) commencing at postnatal day 50. Mice treated with the antisense sequence had significantly extended survival compared to mice injected with a nonsense sequence. Western blot analysis, however, did not reveal a significant reduction in GluR3 protein levels in whole extracts of the lumbar spinal cord. These results suggest that interference with the GluR3 component of the AMPA receptor assembly may be a novel strategy for controlling excitotoxic destruction of motor neurons and may lead to new therapeutic opportunities for the treatment of human ALS. (C) 2004 Wiley-Liss, Inc.