Freese A, Kaplitt M G, O'Connor W M, Abbey M, Langer D, Leone P, O'Connor M J, During M J
Division of Neurosurgery, University of Pennsylvania School of Medicine, Philadelphia 19104-6380, USA.
Epilepsia. 1997 Jul;38(7):759-66. doi: 10.1111/j.1528-1157.1997.tb01462.x.
Virus vectors capable of transferring genetic information into human cells provide hope for improved therapy in several neurological diseases, including epilepsy. We evaluated the ability of an adeno-associated virus (AAV) vector to transfer and cause expression of a lacZ marker gene in brain slices obtained from patients undergoing temporal lobectomy for control of medically intractable seizures.
Human brain slices were injected with an AAV vector (AAVlacZ) encoding Escherichia coli beta-galactosidase and incubated for as long as 24 h. The presence of lacZ mRNA. beta-galactosidase protein and enzymatic activity were assayed by reverse transcriptase polymerase chain reaction (rtPCR), immunocytochemistry, and the X-Gal technique, respectively.
AAVlacZ directed the expression in human epileptogenic brain of E. coli beta-galactosidase that had functional activity. Expression was observed in < or =5 h and was sustained for as long as the slices were viable. Morphological analysis indicated that neurons were preferentially transfected, and there was no evidence of cytotoxicity.
Our results confirm the feasibility of using AAV vectors to transfer genes into the human CNS and in particular, into neurons. Replacement of the lacZ gene with a functional gene modulating hippocampal neuronal physiology, might allow a localized genetic intervention for focal seizures based on the stereotaxic or endovascular delivery of such a vector system into the appropriate brain region.
能够将遗传信息传递到人类细胞中的病毒载体为包括癫痫在内的多种神经疾病的治疗改善带来了希望。我们评估了腺相关病毒（AAV）载体在从因控制药物难治性癫痫而接受颞叶切除术的患者获取的脑切片中传递并导致lacZ标记基因表达的能力。
将编码大肠杆菌β-半乳糖苷酶的AAV载体（AAVlacZ）注入人脑切片，并孵育长达24小时。分别通过逆转录聚合酶链反应（rtPCR）、免疫细胞化学和X-Gal技术检测lacZ mRNA、β-半乳糖苷酶蛋白的存在及酶活性。
AAVlacZ指导了具有功能活性的大肠杆菌β-半乳糖苷酶在人类致痫性脑中的表达。在≤5小时时观察到表达，并且只要切片存活表达就持续存在。形态学分析表明神经元优先被转染，且没有细胞毒性的证据。
我们的结果证实了使用AAV载体将基因传递到人类中枢神经系统，特别是传递到神经元中的可行性。用调节海马神经元生理学的功能基因替换lacZ基因，可能允许基于将这种载体系统立体定向或血管内递送到适当脑区而对局灶性癫痫发作进行局部基因干预。
