TH and GTPCHI gene therapy for Parkinson's disease

描述(由申请人提供)：这项提案汇集了三个经验丰富的小组，提出了在帕金森病(PD)的啮齿动物(Lund)和非人灵长类(Rush)模型中使用腺相关病毒(Gainesville)进行的一系列综合实验。一种共识是，位点特异性重组腺相关病毒介导的纹状体内L-多巴转移可能是治疗帕金森病的有用策略。然而，在需要进行关键的疗效和安全性研究之前，甚至不能开始考虑使用这种方法的临床试验。为此，本申请将探讨三个研究主题。首先，我们将在猴子身上进行以载体优化设计的研究。我们将确定非人灵长类AAV的最佳血清型，并建立TH和GTPCH1的最佳比例。目前，使用的比例为1：1。然而，我们相信，由于GTPCH1的运动，更高的比率将更加有效，因此我们将能够提供更多TH，并最终提供更多LDOPA。这些研究还将提供关键的灵长类“放大”数据，这些数据将与未来的临床试验相关。第二个研究主题是功效。TH/GTPCH1基因传递方法已经被证明在逆转药物诱导的和自发的帕金森病啮齿动物模型中的运动障碍方面是有效的。目前的建议将在MPTP治疗的猴子身上建立疗效，这是目前可用的治疗帕金森病的最佳动物模型。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。我们已经证明了rAAV-LDOPA逆转已经在6-OHDA损伤的大鼠中表现出运动障碍，并有新的数据表明rAAV-LDOPA可以防止运动障碍的出现。在这一新的应用中，我们将评估5-羟色胺在大鼠运动障碍表达中的作用。此外，还将评估“热点”和“广泛”注射TH/GTPCH1对疗效和运动障碍的影响。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，我们将验证这样的假设，即基因传递的左旋多巴既可以逆转已经表现出的异动症，又可以延缓新的异动症的出现。这些研究将确定LDOPA基因传递的安全性和有效性，并确定这种方法是否适合临床试验。

DESCRIPTION (provided by applicant): 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.