Sun Mei, Zhang Guo-Rong, Kong Lingxin, Holmes Courtney, Wang Xiaodan, Zhang Wei, Goldstein David S, Geller Alfred I
Department of Neurology, West Roxbury VA Hospital/Harvard Medical School, 1400 VFW Parkway, West Roxbury, MA 02132, USA.
Hum Gene Ther. 2003 Mar 20;14(5):415-24. doi: 10.1089/104303403321467180.
We previously reported long-term biochemical and behavioral correction of the 6-hydroxydopamine (6-OHDA) rat model of Parkinson's disease (PD) by expression of tyrosine hydroxylase (TH) in the partially denervated striatum, using a herpes simplex virus type 1 (HSV-1) vector. This study had a number of limitations, including the use of a helper virus packaging system, limited long-term expression, and expression of only TH. To address these issues, we developed a helper virus-free packaging system, a modified neurofilament gene promoter that supports long-term expression in forebrain neurons, and a vector that coexpresses TH and aromatic amino acid decarboxylase (AADC). Coexpression of TH and AADC supported high-level (80%), behavioral correction of the 6-OHDA rat model of PD for 5 weeks. Biochemical correction included increases in extracellular dopamine and DOPAC concentrations between 2 and 4 months after gene transfer. Histologic analyses demonstrated neuronal-specific coexpression of TH and AADC at 4 days to 7 months after gene transfer, and cell counts revealed 1000 to 10,000 TH positive cells per rat at 2 months after gene transfer. This improved system efficiently corrects the rat model of PD.
我们之前报道过,利用1型单纯疱疹病毒(HSV-1)载体在部分去神经支配的纹状体中表达酪氨酸羟化酶(TH),可对帕金森病(PD)的6-羟基多巴胺(6-OHDA)大鼠模型进行长期的生化和行为矫正。这项研究存在一些局限性,包括使用辅助病毒包装系统、长期表达受限以及仅表达TH。为了解决这些问题,我们开发了一种无辅助病毒的包装系统、一种经修饰的神经丝基因启动子,该启动子可在前脑神经元中支持长期表达,以及一种能共表达TH和芳香族氨基酸脱羧酶(AADC)的载体。TH和AADC的共表达支持对PD的6-OHDA大鼠模型进行高水平(80%)的行为矫正达5周。生化矫正包括在基因转移后2至4个月细胞外多巴胺和DOPAC浓度增加。组织学分析表明,在基因转移后4天至7个月TH和AADC呈神经元特异性共表达,细胞计数显示在基因转移后2个月每只大鼠有1000至10000个TH阳性细胞。这种改进的系统能有效矫正PD大鼠模型。