Gene therapy targeting striatal dysfunction for Parkinson’s disease

项目摘要 帕金森病（PD）的特征是主要由中脑缺失引起的运动异常 多巴胺（DA）细胞，其显著调节纹状体神经元。因此，DA消耗与改变 纹状体投射神经元（SPNs）。SPN失调通过显著的形态学和病理学改变来证明。 生理变化，如许多离体和体内研究所示。此外，我们对灵长类动物的研究 模型和患者揭示了SPN的病理性活动过度。这种过度活跃的一个关键因素是 兴奋性谷氨酸信号。最近，选择性纹状体阻滞 NMDAR。该阻断可减少多动性并控制对DA替代的改变的神经元反应。 此外，它对帕金森病运动症状有显着影响。因此，数据支持减少 SPN上的NMDAR信号传导可能在PD中具有治疗作用。鉴于大部分保守的结构， 由于NMDAR在大脑区域中的作用，药物治疗通常受到广泛的药物作用的限制。我们 最近测试了靶向纹状体中NMDAR亚基表达的基因疗法。我们的初步数据 在啮齿类动物中进行的探索性试验表明，GluN2亚基的基因敲低（KD）确实可以 为改善PD的运动症状提供了一种新的治疗策略。因此，数据支持高级临床前 在非人灵长类动物（NHP）中进行GluN2基因KD的研究，以广泛评估该基因治疗在 PD的黄金标准模型。 在这个项目中，我们计划证明GluN2基因KD对各种运动和认知症状的“功效”。 的PD。这些研究将使用灵长类MPTP模型和纹状体中的shRNA病毒载体注射来抑制MPTP。 SPN中GluN2的基因表达。一系列的运动和认知测试将在很长一段时间内使用 病毒给药后，以确定稳定的慢性效应。利用NHP中的PD建模， 还将评估不同的疾病阶段。我们还包括额外的结果措施，以评估 靶向纹状体中NMDAR的适当组装的“安全性”。最后，我们将验证这种基因疗法 电生理学数据证明了对改变的SPN活性的特异性作用，并应用光遗传学。 用于NHP记录中的细胞分辨率。总体而言，本提案中的研究策略基于最佳动物 模型，证明有效的病毒载体，对运动，认知和其他行为的广泛测试，以及 用于电生理学的精密工具我们希望这些研究的结果能够证明疗效和安全性 翻译数据支持进一步开发纹状体GluN2基因治疗PD。

Project Summary Parkinson’s disease (PD) is characterized by motor abnormalities primarily caused by loss of midbrain dopamine (DA) cells, which significantly modulate striatal neurons. DA depletion is thus associated with altered function of striatal projection neurons (SPNs). SPN dysregulation is evidenced by significant morphological and physiological changes, as shown in numerous ex-vivo and in-vivo studies. Furthermore, our studies in primate models and patients have revealed pathological hyperactivity of SPNs. A key contributor to this hyperactivity is the excitatory glutamate signaling. This has been recently demonstrated with selective striatal blockade of NMDARs. The block reduces hyperactivity and controls the altered neuronal responses to DA replacement. Furthermore, it has significant effects on parkinsonian motor symptoms. Therefore, data support that reducing NMDAR signaling on SPNs may have therapeutic effects in PD. Given the largely conserved structure of NMDARs across brain regions, pharmacotherapies are generally limited by widespread drug actions. We recently tested gene therapies targeting NMDAR subunit expression in the striatum. Our preliminary data generated with exploratory tests in rodents showed that, indeed, gene knockdown (KD) of GluN2 subunits may offer a novel therapeutic strategy to improve motor symptoms of PD. Thus, data support advanced preclinical studies of GluN2 gene KD in non-human primates (NHP) for extensive evaluation of this gene therapy in the gold-standard model of PD. In this project we plan to demonstrate “efficacy” of GluN2 gene KD for various motor and cognitive symptoms of PD. The studies will use primate MPTP models and shRNA viral vector injections in the striatum to suppress gene expression of GluN2 in SPNs. A battery of motor and cognitive tests will be used over a prolonged term post-virus administration to determine stable, chronic effects. Taking advantage of PD modeling in NHPs, different disease stages will also be evaluated. We also include additional outcome measures to evaluate “safety” of targeting the proper assembly of NMDAR in the striatum. Finally, we will validate this gene therapy with electrophysiology data demonstrating specific effects on altered SPN activity, and applying optogenetics. for cell resolution in NHP recordings. Overall, the research strategy in this proposal is based on optimal animal models, viral vectors of proven efficiency, extensive testing of motor, cognitive, and other behaviors, and precision tools for electrophysiology. We expect that results of these studies demonstrate efficacy and safety with translational data to support further development of striatal GluN2 gene therapy for PD.