Gene therapy targeting striatal dysfunction for Parkinson’s disease

项目摘要 帕金森氏病(PD)的特征是运动异常，主要是由中脑缺失引起的 多巴胺(DA)细胞，它显著调节纹状体神经元。因此，DA耗竭与改变有关 纹状体投射神经元的功能。SPN失调的证据是显著的形态和 生理变化，如许多体外和体内研究所显示的那样。此外，我们对灵长类动物的研究 模型和患者都显示出SPN的病理性多动。这种过度活跃的一个关键因素是 兴奋性谷氨酸信号。这一点最近在选择性纹状体阻断中得到了证实。 NMDAR。该阻滞剂可减少过度活动，并控制对DA替代的改变的神经元反应。 此外，它对帕金森氏症的运动症状有显著的影响。因此，数据支持减少 SPN上的NMDAR信号可能对帕金森病有治疗作用。考虑到基本保守的结构 NMDAR跨越大脑区域，药物治疗通常受到广泛的药物作用的限制。我们 最近测试的针对纹状体NMDAR亚单位表达的基因治疗。我们的初步数据 在啮齿动物中的探索性测试表明，的确，GluN2亚基的基因敲除(Kd)可能 为改善帕金森病的运动症状提供新的治疗策略。因此，数据支持高级临床前研究 非人灵长类(NHP)GluN2基因KD的研究为该基因治疗的广泛评价提供了依据 帕金森病的黄金标准模型。 在这个项目中，我们计划展示GluN2基因KD对各种运动和认知症状的“有效性”。 警局的。这些研究将使用灵长类MPTP模型和纹状体内注射shRNA病毒载体来抑制 谷氨酸氨基转移酶基因在SPN中的表达一组运动和认知测试将在较长的时间内使用 病毒后给药以确定稳定的、慢性的影响。利用NHP中的局部放电建模， 还将对不同的疾病阶段进行评估。我们还包括其他结果衡量标准，以进行评估 以NMDAR在纹状体中的适当组装为靶点的安全性。最后，我们将验证这种基因疗法。 电生理学数据显示对改变的SPN活性的特定影响，并应用光遗传学。 用于NHP记录中的单元分辨率。总体而言，本方案的研究策略是基于最优动物 模型，证实有效的病毒载体，对运动、认知和其他行为的广泛测试，以及 电生理学的精密工具。我们期望这些研究的结果能证明有效性和安全性。 为进一步开展纹状体GluN_2基因治疗帕金森病提供了翻译数据。

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.