AAV-mediated gene therapy for CNS disease correction in feline NPC1 disease

项目总结/摘要 C1型尼曼-匹克病（NPC 1病）是一种以溶酶体储存为特征的遗传性疾病 以及进行性小脑共济失调、痴呆、垂直运动障碍、 核上性凝视麻痹，吞咽困难和早逝目前尚无FDA批准的NPC治疗方法1 疾病重复脑池内（IC）给予2-羟丙基-β-环糊精（HP β CD）， 患有NPC 1疾病的猫预防了小脑共济失调的发作，预防了浦肯野细胞死亡， 小脑皮质和小脑皮质胆固醇和神经节苷脂浓度，并增加生存时间。 这些临床前数据将IC HP β CD推进到临床试验中，并证明了其疗效。 然而，HP β CD必须每两周IC给药一次，持续患者的生命，并导致 进行性剂量限制性耳毒性，突出了对这些更小侵入性和更安全的治疗的明确需求 患者 我们假设，使用AAV 9载体将NPC 1递送至大脑的IC基因治疗的优化将使NPC 1的表达水平降低。 有效预防NPC 1疾病相关小脑共济失调和浦肯野细胞病理学， 终身注射且无耳毒性。我们还假设，颈动脉（IV）给药的新的 猫的AAV血清型可以将NPC 1递送到基底神经节和脑干，这些区域未被IC处理 施用HP β CD或AAV 9，并导致肌张力障碍、垂直核上性凝视麻痹，和 吞咽困难因此，在所提出的研究中，我们将评估优化AAV 9介导的免疫调节的方法。 转导最大数量的浦肯野细胞（目的1），评估AAV 9-NPC 1的功效 给药以治疗NPC 1相关小脑疾病的临床、生物化学和组织学方面（目的 2），并评估颈动脉内施用AAV-NPC 1治疗小脑外区域的功效 负责痴呆（大脑皮层），肌张力障碍（基底神经节），和垂直核上性凝视麻痹， 吞咽困难（脑干）（目的3）。 这些概念验证研究将是第一个优化非扩散性膜结合型药物递送的研究。 蛋白质，从而推进基因治疗的许多其他遗传性疾病影响浦肯野细胞 包括脊髓小脑共济失调。而且，这些研究将首次开发出一次性疗法 对于NPC 1疾病，治疗小脑和小脑外疾病，并且不导致耳毒性。

Project Summary/Abstract Niemann-Pick disease type C1 (NPC1 disease) is a hereditary disorder characterized by the lysosomal storage of cholesterol and sphingolipids, and clinical signs of progressive cerebellar ataxia, dementia, vertical supranuclear gaze palsy, dysphagia, and early death. There are no FDA-approved therapies for NPC1 disease. Repeated intracisterna magna (IC) administration of 2-hydroxypropyl-beta-cyclodextrin (HPßCD) in cats with NPC1 disease prevented the onset of cerebellar ataxia, prevented Purkinje cell death, normalized cerebellocortical and cerebrocortical cholesterol and gangliosides concentrations, and increased survival time. These preclinical data advanced IC HPßCD into clinical trials where efficacy has been demonstrated. However, HPßCD must be administered IC every two weeks for the duration of the patient’s life and results in progressive dose-limiting ototoxicity, highlighting a clear need for less invasive and safer therapies for these patients. We hypothesize that optimization of IC gene therapy using an AAV9 vector to deliver NPC1 to the brain will effectively prevent NPC1 disease-associated cerebellar ataxia and Purkinje cell pathology without repeated lifelong injections and without ototoxicity. We also hypothesize that intracarotid (IV) administration of a novel AAV serotype to cats can deliver NPC1 to the basal ganglia and brainstem, regions which are untreated by IC administration of HPßCD or AAV9, and are responsible for dystonia, vertical supranuclear gaze palsy, and dysphagia. Therefore, in the proposed studies we will assess methods to optimize AAV9-mediated transduction of the greatest number of Purkinje cells (Aim 1), evaluate the efficacy of AAV9-NPC1 administration to treat clinical, biochemical, and histologic aspects of NPC1-associated cerebellar disease (Aim 2), and evaluate the efficacy of intracarotid AAV-NPC1 administration to treat extracerebellar regions responsible for dementia (cerebral cortex), dystonia (basal ganglia), and vertical supranuclear gaze palsy and dysphagia (brainstem) (Aim 3). These proof-of-concept studies will be the first to optimize the delivery of a non-diffusible membrane-bound protein to Purkinje cells, thereby advancing gene therapy for many other genetic diseases affecting Purkinje cells including spinocerebellar ataxias. Moreover, these studies will be the first to develop a one-time therapy for NPC1 disease that treats both cerebellar and extracerebellar disease and results in no ototoxicity.