Viral-mediated temporally controlled dopamine production in a rat model of Parkinson disease
Viral-mediated temporally controlled dopamine production in a rat model of Parkinson disease
复制标题
DOI：
10.1016/j.ymthe.2005.08.009
复制
发表时间：
2006-01-01
影响因子：
12.4
通讯作者：
Muramatsu, S
中科院分区：
文献类型：
--
作者：
Li, XG;Okada, T;Muramatsu, S
为了避免转基因产物过度合成可能导致的基因治疗的不良影响，有必要对基因表达进行调控。为了减少转基因表达，我们开发了一种利用可诱导的Cre重组酶的病毒载体介导的体细胞调控系统。将表达Cre重组酶的重组腺相关病毒(AAV)载体与表达多巴胺合成酶的重组腺相关病毒载体(AAV)一起注射给帕金森病模型大鼠。用人工合成的雌激素受体调节剂4-羟基塔诺昔芬处理，激活了转导神经元内的Cre重组酶，并诱导了IoxP位点两侧的酪氨酸羟基酶(TH)编码序列的选择性切除，导致转基因介导的多巴胺合成减少。利用这一策略，保留了芳香族L氨基酸脱羧酶(AADC)的活性，以便AADC的底物(L)-3，4-二羟基苯丙氨酸(L-多巴)能够在纹状体内转化为多巴胺，并保留(L)-多巴的治疗作用，即使在多巴胺过度产生的情况下TH表达减少也是如此。我们的数据表明，病毒载体介导的可诱导的Cre重组酶可以作为体内的分子开关，允许对转基因表达进行时空控制，从而潜在地增加基因治疗的安全性。
Regulation of gene expression is necessary to avoid possible adverse effects of gene therapy due to excess synthesis of transgene products. To reduce transgene expression, we developed a viral vector-mediated somatic regulation system using inducible Cre recombinase. A recombinant adeno-associated virus (AAV) vector expressing Cre recombinase fused to a mutated ligand-binding domain of the estrogen receptor alpha (CreER(T2)) was delivered along with AAV vectors expressing dopamine-synthesizing enzymes to rats of a Parkinson disease model. Treatment with 4-hydroxytarnoxifen, a synthetic estrogen receptor modulator, activated Cre recombinase within the transduced neurons and induced selective excision of the tyrosine hydroxylase (TH) coding sequence flanked by IoxP sites, leading to a reduction in transgene-mediated dopamine synthesis. Using this strategy, aromatic L-amino acid decarboxylase (AADC) activity was retained so that (L)-3, 4-dihydroxyphenylalanine (L-dopa), a substrate for AADC, could be converted to dopamine in the striatum and the therapeutic effects of (L)-dopa preserved, even after reduction of TH expression in the case of dopamine overproduction. Our data demonstrate that viral vector-mediated inducible Cre recombinase can serve as an in vivo molecular switch, allowing spatial and temporal control of transgene expression, thereby potentially increasing the safety of gene therapy.