Adeno-associated virus-mediated expression and constitutive secretion of galanin suppresses limbic seizure activity in vivo
Adeno-associated virus-mediated expression and constitutive secretion of galanin suppresses limbic seizure activity in vivo
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DOI：
10.1016/j.ymthe.2006.04.004
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发表时间：
2006-07-01
影响因子：
12.4
通讯作者：
McCown, Thomas J.
中科院分区：
文献类型：
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作者：
McCown, Thomas J.
难治性颞叶癫痫是基因治疗的理想靶点，但治疗的成功取决于抑制边缘系统癫痫发作的能力。构建腺相关病毒载体（AAV），其中纤连蛋白分泌信号序列（FIB）位于甘丙肽（AAV-FIB-GAL）或绿色荧光蛋白（AAV-FIB-GFP）的编码序列之前，构建体表达并组成性分泌基因产物。双侧AAV-FIB-GAL输注到大鼠梨状皮质（2 μ l/侧）中显著减弱了红藻氨酸诱导的癫痫发作（10 mg/kg，ip），使得11/12只大鼠没有表现出边缘癫痫发作，而剩余的大鼠仅表现出短暂的单次III类癫痫发作。这种AAV-FIB-GAL输注也防止了电图癫痫发作活动。相反，双侧AAV-FIB-GFP输注不改变行为或电图癫痫发作活动。由于先前的癫痫发作暴露可能影响载体功效，另一组大鼠每天接受梨状皮质的电刺激，直到引起连续三次V类癫痫发作。随后，将AAV-FIB-GAL或AAV-FIB-GFP（3 μ l/30分钟）注入电极区域。一周后，AAV-FIB-GAL大鼠表现出引起边缘癫痫发作活动所需的刺激电流的显著增加，而AAV-FIB-GFP没有改变癫痫发作阈值。因此，AAV介导的甘丙肽表达和分泌显著抑制体内边缘癫痫发作活性。
Intractable temporal lobe epilepsy presents an ideal target for gene therapy, but therapeutic success depends upon the ability to suppress limbic seizure activity. Adeno-associated virus vectors (AAV) were constructed in which the fibronectin secretory signal sequence (FIB) preceded the coding sequence for galanin (AAV-FIB-GAL) or green fluorescent protein (AAV-FIB-GFP), constructs that express and constitutively secrete the gene product. Bilateral AAV-FIB-GAL infusion into the rat piriform cortex (2 [mu l/side) significantly attenuated kainic acid-induced seizures (10 mg/kg, ip) such that 11/12 rats exhibited no limbic seizures, while the remaining rat exhibited only a brief, single class III seizure. This AAV-FIB-GAL infusion also prevented electrographic seizure activity. In contrast, bilateral AAV-FIB-GFP infusion did not alter either behavioral or electrographic seizure activity. Since prior seizure exposure could influence vector efficacy, another group of rats received daily electrical stimulation of the piriform cortex until three consecutive class V seizures were elicited. Subsequently, AAV-FIB-GAL or AAV-FIB-GFP (3 mu l/30 min) was infused into the area of the electrode. One week later the AAV-FIB-GAL rats exhibited a significant increase in the stimulation current necessary to evoke limbic seizure activity, while AAV-FIB-GFP did not alter the seizure threshold. Thus, AAV-mediated galanin expression and secretion significantly suppress limbic seizure activity in vivo.