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

项目摘要/摘要 Niemann-Pick病C1型(NPC1病)是一种以溶酶体储存为特征的遗传性疾病 胆固醇和鞘脂，以及进行性小脑性共济失调、痴呆、垂直性 核上性凝视麻痹、吞咽困难和早死。目前还没有FDA批准的NPC1疗法 疾病。枕大池内重复注射2-羟丙基-β-环糊精(HP？CD) 患有NPC1病的猫防止了小脑性共济失调的发作，防止了浦肯野细胞的死亡，正常化 小脑皮质和大脑皮层胆固醇和神经节苷脂浓度增加，存活时间延长。 这些临床前数据将IC HPüCD推进到已证明有效的临床试验中。 然而，在患者生命的持续时间内，HP？CD必须每两周给药一次，结果是 渐进性剂量限制耳毒性，突出了对这些疾病进行侵入性更小、更安全的治疗的明显需要 病人。 我们假设，使用AAV9载体将NPC1运送到大脑的IC基因治疗的优化将 有效预防NPC1病相关性小脑性共济失调和浦肯野细胞病变 终身注射，没有耳毒性。我们还假设颈动脉内(IV)给药一种新的 猫的AAV血清型可将NPC1传递到未经IC治疗的基底节和脑干 给予HP？CD或AAV9，可导致肌张力障碍、垂直核上凝视麻痹和 吞咽困难。因此，在拟议的研究中，我们将评估优化AAV9介导的方法 转导最多的浦肯野细胞(目标1)，评价AAV9-NPC1的疗效 给药治疗NPC1相关性小脑疾病的临床、生化和组织学方面(目的 2)，并评价颈动脉注射AAV-NPC1治疗小脑外区的疗效。 负责痴呆(大脑皮层)、肌张力障碍(基底节)和垂直核上凝视麻痹和 吞咽困难(脑干)(目标3)。 这些概念验证研究将是第一次优化非扩散膜结合的输送 蛋白质转移到浦肯野细胞，从而推进了影响浦肯野的许多其他遗传性疾病的基因治疗 包括脊髓小脑性共济失调在内的细胞。此外，这些研究将是第一次开发一次性疗法。 对于NPC1疾病，既治疗小脑疾病又治疗小脑外疾病，并且没有耳毒性。

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.