Studies on striatal function using adeno-associated viral vectors

使用腺相关病毒载体研究纹状体功能

基本信息

批准号：
17590890
负责人：
MURAMATSU Shin-ichi
金额：
$ 2.24万
依托单位：
Jichi Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590890/
关键词：
striatum dopamine putamen gene therapy PET adeno-associated virus Parkinson disease cynomolgus monkey

项目摘要

Recombinant adeno-associated virus (AAV) vector-mediated gene transfer of tyrosine hydroxylase (TH), aromatic L-amino acid decarboxylase (AADC), and guanosine triphosphate cyclohydrolase 1 (GCH), which are dopamine (DA)-synthesizing enzymes, provided behavioral recovery in a primate model of Parkinson's disease (PD) with efficient and long-term transduction of striatal neurons. We applied positron emission tomography (PET) to assess trangene-mediated DA synthesis in vivo in a primate model of PD. Cynomolgus macaques (Macaca fascicularis) were chronically treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to make bilateral striatal lesions. After the animals had been behaviorally stable for at least two months, mixtures of AAV vectors expressing the DA-synthesizing enzymes were injected into the unilateral putamen stereotaxically. Use of [^<11>C], a short half-life (20 min) tracer, has made it possible to obtain sequential imaging data, not only during the early steps of D … More A metabolism, but also during DA receptor binding and DA transporter availability in the same subject on the same day. In the AAV vector-injected putamen, remarkable and persistent increase of [^<11>C]L-dopa uptake was observed, reflecting restoration of AADC activity, while bindings of [^<11>C]2_-carbomethoxy-3_(4-fluorophenyl)tropane (_-CFT), DA transporter ligand, and [^<11>C] raclopride, a selective D_2-like receptor antagonist, remained unchanged. DA receptor availability to [^<11>C] raclopride was significantly reduced in the AAV vector-injected putamen following L-dopa administration, suggesting additional synthesis of DA in response to the exogenous L-dopa.A recombinant adeno-associated virus (AAV) vector that expressing Cre recombinase fused to a mutated ligand binding domain of estrogen receptor _(CreER^<T2>) was delivered along with AAV vectors that expressing dopamine-synthesizing enzymes to rats with a model of Parkinson's disease. Treatment with 4-hydroxytamoxifen, a synthetic estrogen receptor modulator, activated the Cre recombinase in the transduced neurons and induced selective excision of the tyrosine hydroxylase (TH) coding sequence that was flanked by loxP sites, leading to the reduction of transgene-mediated dopamine synthesis. Less

重组腺癌相关病毒（AAV）载体介导的基因基因触动羟化酶（Th）和三磷酸三磷酸鸟嘌呤环己酶1（GCH），这些酶是多巴胺（DA） - 结合酶，在帕克森病（Parkinson's Parkinson's Diseation's Diseation's Parkinson's Plackinson's Diseation（Pd）中提供的行为恢复，明天（PET）的效率和转导，在PD的灵长类动物模型中评估体内的trangene介导的DA合成。 2,3,6-四氢吡啶（MPTP）使动物在行为上稳定至少两个月11> c]使得获得顺序成像数据，不仅在D的早期步骤中……在同一天，同一受试者中的DA受体结合和DA转运蛋白的可用性。观察到[^<11> c] l-dopa摄取的摄取，反映了AADC活性的恢复，而[^<11> c] 2_--核甲氧替-3_（4-氟苯基）的结合是转运蛋白配体，而[^<11 > c] raclopride，一种选择性的D_2样受体拮抗剂，DA受体的可用性在L-DOPA给药后，对[^<11> c] AAV向量注射的鬼phiment型，表明DA对da的适当合成响应于外源L-Dopa.A重组腺相关的病毒（AAV）载体表达R _（Creer^<t2>）的载体与AAV载体一起传递，以用帕金森模型表达多巴胺合成的酶。）编码序列是由LOXP位点重新介导的转基因介导的多巴胺合成的序列。