Gene Therapy and Seizures

基因治疗和癫痫发作

• 批准号: 6577150
• 负责人: Thomas J. McCown
• 金额: $ 30.02万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6577150
• 关键词: adeno associated virus group; cell death; epilepsy; fibronectins; galanin; gene expression; gene therapy; genetic transduction; hippocampus; histochemistry /cytochemistry; inferior colliculus; inhibitor /antagonist; kainate; laboratory rat; nervous system disorder therapy; neuropeptides; nonhuman therapy evaluation; partial seizure; pilocarpine; protein biosynthesis; transfection /expression vector

DESCRIPTION (provided by applicant): Theoretically, viral vector gene therapy holds great promise for the treatment of epilepsy, where in vivo expression of foreign genes could potentially suppress focal seizure sensitivity. Neurotransmitter receptors and ion channels offer obvious gene therapy targets, but a successful outcome is dependent upon the pattern of viral Vector transduction which to date cannot be predicted a priori. One means to circumvent this potential impediment would be to express an endogenous inhibitory neuropeptide that is subsequently secreted from the transduced cell. Two endogenous peptides, galanin (GAL) and neuropeptide Y (NPY), both can potently suppress limbic seizure activity. Recent preliminary findings with adeno-associated virus vectors (AAV) show that the secretion signal sequence for the laminar protein, fibronectin, can secrete vector-derived gene product in vitro and when placed in front of the GAL coding sequence, suppresses in vivo focal seizure sensitivity in a regulatable fashion and attenuates kainic acid-induced cell death in the hippocampus. Therefore, the present proposal will test the hypothesis that in vivo secretion of vector derived GAL or NPY will suppress acute seizure activity, reduce kainic acid seizure associated cell death and retard the spontaneous seizure activity that develops after pilocarpine administration. First, recombinant AAV vectors containing fibronectin secretory sequence in front of GAL or NPY coding sequences will be infused into the inferior colliculus, and both the persistence of seizure suppression and the ability to regulate gene expression will be evaluated. Then, these same recombinant AAV vectors will be tested in the kainic acid seizure model, evaluating if transduction of hippocampal hilar neurons can alter acute seizure sensitivity and seizure-induced cell damage. Subsequently, it will be determined if these AAV vectors can prevent the development of spontaneous seizures following pilocarpine-induced seizures. The findings from these studies could lead to a novel gene therapeutic approach to the treatment of epilepsy.

描述（由申请人提供）：理论上，病毒载体基因疗法对于治疗癫痫有很大希望，其中外源基因的体内表达可能会抑制局灶性癫痫敏感性。神经递质受体和离子通道提供了明显的基因治疗靶标，但成功的结果取决于病毒载体转导的模式，而迄今为止尚无法先验预测该模式。规避这种潜在障碍的一种方法是表达随后从转导细胞中分泌的内源抑制性神经肽。两种内源性肽，甘丙肽 (GAL) 和神经肽 Y (NPY)，都可以有效抑制边缘系统癫痫发作活动。最近对腺相关病毒载体 (AAV) 的初步研究结果表明，层状蛋白纤连蛋白的分泌信号序列可以在体外分泌载体衍生的基因产物，并且当放置在 GAL 编码序列前面时，可以抑制体内局灶性癫痫发作以可调节的方式调节敏感性并减弱红藻氨酸诱导的海马细胞死亡。因此，本提案将检验以下假设：载体衍生的 GAL 或 NPY 的体内分泌将抑制急性癫痫发作活动，减少红藻氨酸癫痫发作相关的细胞死亡，并延缓毛果芸香碱给药后发生的自发癫痫发作活动。首先，将在GAL或NPY编码序列前面含有纤连蛋白分泌序列的重组AAV载体注入下丘，评估癫痫发作抑制的持续性和调节基因表达的能力。然后，这些相同的重组 AAV 载体将在红藻氨酸癫痫模型中进行测试，评估海马门神经元的转导是否可以改变急性癫痫敏感性和癫痫引起的细胞损伤。随后，将确定这些 AAV 载体是否可以预防毛果芸香碱诱导的癫痫发作后自发性癫痫发作的发展。这些研究的结果可能会带来一种治疗癫痫的新基因治疗方法。