Theoretically, gene therapy techniques offer an attractive alternative treatment option for intractable, focal epilepsies. Although logical gene therapy targets include excitatory and inhibitory receptors, variable viral vector tropism interjects an uncertainty as to the direction of change, seizure suppression, or seizure sensitization. To circumvent this therapeutic liability, adeno-associated virus (AAV) vectors have been constructed where the gene product is constitutively secreted from the transduced cell. Using AAV vectors, the fibronectin secretory signal sequence (FIB) was placed in front of the coding sequence for green fluorescent protein or the active portion of the neuroactive peptide galanin (GAL). Subsequent studies showed that these vectors supported expression and constitutive secretion of these gene products from transfected cells in vitro. More importantly, upon transduction in vivo, AAV-FIB-GAL vectors significantly attenuated focal seizure sensitivity, and this seizure attenuation could be controlled in vivo by using a tetracyclineregulated promoter. The expression and constitutive secretion of green fluorescent protein, or the expression of GAL alone, exerted no effect on focal seizure sensitivity. Moreover, unilateral infusion of the AAV-FIB-GAL vectors into the hippocampus prevented kainic acid-induced hilar cell death. With regard to limbic seizures, bilateral infusion of AAV-FIB-GAL vectors into the piriform cortex prevented both behavioral and localized electrographic seizure activity after the peripheral administration of kainic acid. Also, when rats were electrically kindled to class V seizure activity, subsequent infusion of AAV-FIB-GAL proved capable of significantly elevating the seizure initiation threshold. Thus, these studies clearly demonstrate the anti-seizure effectiveness of AAV vector-mediated expression and constitutive secretion of galanin.

中文翻译：

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腺相关病毒载体介导的甘丙肽表达和组成型分泌抑制边缘癫痫发作活动。

从理论上讲，基因治疗技术为顽固性局灶性癫痫提供了一种有吸引力的替代治疗选择。虽然合乎逻辑的基因治疗目标包括兴奋性和抑制性受体，但可变的病毒载体趋向性在变化方向、癫痫抑制或癫痫致敏方面存在不确定性。为了规避这种治疗责任，已经构建了腺相关病毒 (AAV) 载体，其中基因产物从转导细胞中组成性分泌。使用 AAV 载体，将纤连蛋白分泌信号序列 (FIB) 置于绿色荧光蛋白的编码序列或神经活性肽甘丙肽 (GAL) 的活性部分之前。随后的研究表明，这些载体支持这些基因产物从转染细胞中的表达和组成型分泌体外。更重要的是，在体内转导后，AAV-FIB-GAL 载体显着减弱了局灶性癫痫发作的敏感性，并且可以在体内控制这种癫痫发作的减弱通过使用四环素调节的启动子。绿色荧光蛋白的表达和组成型分泌，或单独的 GAL 表达，对局灶性癫痫敏感性没有影响。此外，将 AAV-FIB-GAL 载体单侧注入海马可防止红藻氨酸诱导的肺门细胞死亡。关于边缘癫痫发作，在外周施用红藻氨酸后，将 AAV-FIB-GAL 载体双侧注入梨状皮层可防止行为和局部电图癫痫活动。此外，当大鼠被电激发到 V 类癫痫活动时，随后注入 AAV-FIB-GAL 证明能够显着提高癫痫发作起始阈值。因此，