TH and GTPCHI gene therapy for Parkinson's disease

这项建议汇集了三个经验丰富的小组，提出了一系列集成的实验，使用 帕金森病啮齿动物(Lund)和非人灵长类(Rush)模型中的腺相关病毒(Gainesville) 疾病(PD)。有一种观点认为，位点特异性重组腺相关病毒介导的纹状体L-多巴传递可能是一种 治疗帕金森病的有效策略。然而，使用这种方法的临床试验甚至不能开始 在需要进行关键疗效和安全性研究之前进行考虑。为此，三个 研究主题将在本申请中进行探索。首先，我们将在猴子身上进行研究， 向量优化。我们将确定非人灵长类动物的最佳AAV血清型，并建立 TH与GTPCH1的最佳比例目前采用1：1的比例。然而，我们相信，更高的比率将 由于GTPCH1的推动，我们将更加有效，因此我们将能够交付更多TH，并最终 更多的LDOPA。这些研究还将在灵长类动物中提供关键的“放大”数据，这将与 未来的临床试验。第二个研究主题是功效。TH/GTPCH1基因传递方法具有 已经被证明在逆转药物引起的啮齿动物自发运动障碍方面是有效的 帕金森病的模型。目前的建议将在MPTP治疗的猴子身上建立疗效，这是最好的动物模型 可用于PD。第三个目标是安全。关于功能安全，我们主要关注的是运动障碍。 我们已经证明，rAAV-LDOPA逆转已经在6-OHDA损害中表现出运动障碍 并有新的数据表明，rAAV-LDOPA可以防止运动障碍的出现。在这件事上 在新的应用中，我们将评估5-羟色胺在大鼠运动障碍表达中的作用。此外， “热点”与“广泛”注射TH/GTPCH1对疗效和运动障碍的影响将是 已评估。运动障碍的猴子模型被认为是研究运动障碍的最佳模型。 在目前的研究中，我们将检验这样一种假设，即基因传递的左旋多巴已经可以逆转 表现出伊多巴运动障碍，延缓新运动障碍的出现。这些研究将确定 LDOPA基因传递的安全性和有效性，并确定该方法是否适合于 临床试验。

This proposal brings together three experienced groups proposing an integrated series of experiments using adeno-associated virus (Gainesville) in rodent (Lund) and nonhuman primate (Rush) models of Parkinson's disease (PD). A concensus is emerging that site-specific rAAV-mediated striatal L-dopa delivery might be a useful strategy for treating PD. However, clinical trials using this approach cannot even begin to be considered before critical efficacy and safety studies need to be performed. Towards this end, three research themes will be explored in this application. First we will perform studies in monkeys designed at vector optimization. We will determine the optimal AAV serotype in nonhuman primates and establish the optimal ratio of TH to GTPCH1.Currently, a 1:1 ratio is utilized. However, we believe that a higher ratio will be more effective due to the kinestics of GTPCH1 and thus we will be able to deliver more TH, and ultimately more LDOPA. These studies will also provide critical "scaling-up" data in primates that will be relevant for futures clinical trials. The second research theme is efficacy. The TH/GTPCH1 gene delivery approach has already been shown to be effective in reversing drug-induced and spontaneous motor deficits in rodent models of PD. The present proposal will establish efficacy in MPTP treated monkeys, the best animal model available for PD. The third aim is safety. With regards to functional safety, our main concern is dyskinesias. We have already demonstrated that rAAV-Ldopa reverses already manifest dyskinesias in 6-OHDA lesioned rats and have new data demonstrating that rAAV-LDOPA prevents the emergence of dyskinesias. IN this new application, we will evaluate the role of serotonin in the expression of dyskinesias in rats. Further, the effect of "hot spot" versus "widespread" delivery of TH/GTPCH1 upon efficacy and dyskinesias will be evaluated. The monkey model of dyskinesias is recognized as the best available for the study of dyskinesias. In the present study we will test the hypothesis that gene delivered levodopa can both reverse already manifest Idopa dyskinesias and delay the emergence of new dyskinesias. These studies will determine the safety and efficacy of gene delivery of LDOPA and determine whether this approach is appropriate for clinical trials.