Adeno-associated virus vector-mediated expression and constitutive secretion of galanin suppresses limbic seizure activity.
Adeno-associated virus vector-mediated expression and constitutive secretion of galanin suppresses limbic seizure activity.
复制标题
DOI：
10.1016/j.nurt.2009.01.004
复制
发表时间：
2009-04
期刊：
影响因子：
--
通讯作者：
McCown TJ
中科院分区：
文献类型：
--
作者：
McCown TJ
从理论上讲，基因治疗技术为难治性局灶性癫痫提供了一种有吸引力的替代治疗选择。虽然合理的基因治疗靶点包括兴奋性和抑制性受体，但可变的病毒载体嗜性干扰了变化方向、癫痫发作抑制或癫痫发作致敏的不确定性。为了避免这种治疗倾向，已经构建了腺相关病毒（AAV）载体，其中基因产物从转导的细胞组成型分泌。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在体外从转染细胞中表达和组成性分泌。更重要的是，在体内转导后，AAV-FIB-GAL载体显著减弱局灶性癫痫发作敏感性，并且这种癫痫发作减弱可以通过使用四环素调节的启动子在体内控制。绿色荧光蛋白的表达和组成性分泌，或GAL单独表达，对局灶性癫痫发作敏感性没有影响。此外，单侧输注AAV-FIB-GAL载体到海马防止红藻氨酸诱导的门细胞死亡。关于边缘癫痫发作，在外周施用红藻氨酸后，将AAV-FIB-GAL载体双侧输注到梨状皮质中防止了行为和局部电图癫痫发作活动。此外，当大鼠被电点燃至V类癫痫发作活动时，随后的AAV-FIB-GAL输注被证明能够显著提高癫痫发作起始阈值。因此，这些研究清楚地证明了腺相关病毒载体介导的甘丙肽表达和组成性分泌的抗癫痫发作有效性。
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 tetracycline-regulated 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.
登录
查看更多内容
影响因子：
12.4
作者：
Haberman, RP;Criswell, HE;McCown, TJ
通讯作者：
McCown, TJ
影响因子：
2.9
作者：
McCown, TJ;Xiao, X;Samulski, RJ
通讯作者：
Samulski, RJ
DOI：
10.1073/pnas.93.21.11382
发表时间：
1996-10-15
影响因子：
11.1
作者：
Naldini, L;Blomer, U;Verma, IM
通讯作者：
Verma, IM
影响因子：
12.4
作者：
McCown, Thomas J.
通讯作者：
McCown, Thomas J.
影响因子：
56.9
作者：
MONYER, H;SPRENGEL, R;SEEBURG, PH
通讯作者：
SEEBURG, PH