Genetic Silencing of Striatal CaV1.3 Calcium Channels as a Potent Antidyskinetic Therapy for PD

纹状体 CaV1.3 钙通道的基因沉默作为 PD 的有效抗运动障碍疗法

• 批准号: 10179502
• 负责人: Jeffrey H Kordower
• 金额: $ 56.84万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10179502
• 关键词: Antiparkinson Agents; Behavior; Brain; Calcium Channel; Cardiovascular system; Chronic; Clinical; Corpus striatum structure; Data; Dendritic Spines; Development; Disease model; Dopamine; Dose; Drug Antagonism; Dyskinetic syndrome; FDA approved; Gene Delivery; Gene Silencing; Genes; Genetic; Gold; Holidays; Individual; Intervention; Involuntary Movements; L-DOPA induced dyskinesia; L-Type Calcium Channels; Levodopa; Link; Macaca; Macaca fascicularis; Mediating; Medical; Messenger RNA; Modeling; Molecular; Motor; NR4A2 gene; Neurons; Parkinson Disease; Parkinsonian Disorders; Pathology; Patients; Pharmaceutical Preparations; Pharmacology; Pharmacotherapy; Phenotype; Preclinical Testing; Prevention; Prevention approach; Primates; Prior Therapy; Rattus; Recombinant adeno-associated virus (rAAV); Reporting; Research; Safety; Series; Severities; Specificity; Synapses; Testing; Therapeutic Intervention; Time; Translating; Validation; Work; clinical application; clinical development; clinically relevant; drug withdrawal; gene therapy; genetic approach; illness length; indexing; interest; nonhuman primate; parkinsonian rodent; pre-clinical; prevent; putamen; response; side effect; small hairpin RNA; statistics; transcription factor; translational study

Project Summary One often debilitating side-effect of standard pharmacotherapy for Parkinson's disease (PD), levodopa administration, are unwanted involuntary movements known as levodopa-induced dyskinesia (LID). Eliminating LID remains a significant unmet need in PD therapy. There are currently no FDA approved drug treatments for LID, yet up to 90% of individuals with PD develop this side-effect. The L-type calcium channel CaV1.3 is a target of interest for LID prevention. Loss of striatal dopamine (DA) in PD results in dysregulation and overactivity of striatal CaV1.3 channels leading to synaptic pathology, including the loss of dendritic spines on striatal spiny projection neurons that appears to be involved in LID. While initial studies delivering pharmacological CaV1.3 channel antagonists reduced LID dose-dependently, the effects were partial and transient, with potential liability for cardiovascular side-effects due to the lack of specificity of existing drugs for the CaV1.3 channel. To provide unequivocal target validation, free of pharmacological limitations, we developed a rAAV-CaV1.3-shRNA to provide continuous, high potency, target-selective, mRNA-level silencing of striatal CaV1.3 channels. We examined whether genetic silencing of these dysregulated calcium channels could prevent LID induction in previously levodopa naïve parkinsonian rats and/or whether it could reverse these abnormal behaviors in parkinsonian rats already expressing a severe LID phenotype. In our `LID prevention studies' we found that gene level silencing of striatal CaV1.3 channels in severely parkinsonian rats, prior to the introduction of levodopa provides uniform and complete protection against the induction of LID, and that the antidyskinetic benefit is sustained over time even with high doses of daily levodopa. In our `LID reversal studies' we observed that rAAV-mediated CaV1.3 silencing in parkinsonian rats with already established LID could ameliorate these behaviors, with a one-week drug withdrawal 'drug holiday' appearing to be beneficial and/or necessary. Importantly this approach did NOT interfere with motor benefit of levodopa and showed a tendency to enhance motoric response to low dose levodopa. Gene delivery resulting in striatal CaV1.3 silencing provides some of the most profound antidyskinetic benefit reported to date. If these findings can be translated into a clinical application with a similar magnitude, this would provide a much-needed breakthrough in treatment of individuals with PD and would allow the most powerful antiparkinsonian therapy ever identified to work unabated through the duration of the disease. In the current application we propose a series of translational studies in rats and nonhuman primates that will allow us to expand upon these initial proof-of-principle studies and test specific hypotheses of safety and efficacy that will be required for the clinical development of genetic silencing of striatal CaV1.3 channels for LID.

项目概要 左旋多巴是帕金森病 (PD) 标准药物治疗的一种常见副作用 服用药物后，会产生不必要的不​​自主运动，称为左旋多巴引起的运动障碍（LID）。消除 LID 仍然是 PD 治疗中一个未满足的重大需求。目前尚无 FDA 批准的药物治疗方法 LID，但高达 90% 的 PD 患者会出现这种副作用。 L 型钙通道 CaV1.3 是 LID 预防的兴趣目标。 PD 中纹状体多巴胺 (DA) 的丧失会导致失调和 纹状体 CaV1.3 通道过度活跃，导致突触病理，包括树突棘缺失 纹状体多刺投射神经元似乎与 LID 有关。虽然初步研究成果 药理学 CaV1.3 通道拮抗剂以剂量依赖性方式降低 LID，效果是部分的且 暂时性的，由于现有药物缺乏特异性，可能会产生心血管副作用 CaV1.3 通道。为了提供明确的目标验证，不受药理学限制，我们 开发了 rAAV-CaV1.3-shRNA，以提供连续、高效、靶点选择性、mRNA 水平的沉默 纹状体 CaV1.3 通道。我们检查了这些失调的钙通道是否存在基因沉默 可以预防先前未接受过左旋多巴治疗的帕金森病大鼠的 LID 诱导和/或是否可以逆转 帕金森病大鼠的这些异常行为已经表现出严重的 LID 表型。在我们的“LID”中 预防研究中，我们发现严重帕金森病大鼠纹状体 CaV1.3 通道的基因水平沉默， 在引入左旋多巴之前提供均匀和完整的保护，防止 LID 的诱导，并且 即使每天服用高剂量的左旋多巴，抗运动障碍的效果也会随着时间的推移而持续。在我们的“LID”中 逆转研究中，我们观察到 rAAV 介导的 CaV1.3 在帕金森病大鼠中的沉默已经 建立 LID 可以改善这些行为，为期一周的戒毒“药物假期”似乎可以改善这些行为。 是有益的和/或必要的。重要的是，这种方法不会干扰左旋多巴的运动益处，并且 显示出增强对低剂量左旋多巴的运动反应的趋势。基因传递导致纹状体 CaV1.3 沉默提供了迄今为止报道的一些最深远的抗运动障碍益处。如果这些发现 可以转化为类似规模的临床应用，这将提供急需的 帕金森病患者治疗的突破，将带来最有效的抗帕金森病治疗 曾被确定在疾病期间其工作有增无减。在当前的应用中，我们提出了一个 在大鼠和非人类灵长类动物中进行的一系列转化研究将使我们能够扩展这些最初的研究 原理验证研究并测试临床所需的安全性和有效性的具体假设 开发用于 LID 的纹状体 CaV1.3 通道基因沉默。