Novel Neuronal Transport Mechanisms of Lyosomal Enzymes

描述（由申请人提供）：由于脑储存物质的积累而导致的认知缺陷可能是与溶酶体储存病相关的最严重的临床症状。中枢神经系统的缺陷也很难治疗。我们最近将AAV载体直接注射到VII型粘多糖病（MPS VII）小鼠模型的玻璃体中。在视网膜和视神经中观察到高水平的GUSB活动。我们还在NTS VII小鼠的大脑中发现了GUSB的活性。GUSB活动主要在视束中检测到，特别是外侧膝状丘和上丘。有趣的是，溶酶体储存物质的减少超出了视觉系统，并进入海马和大脑皮层。因此，这种方法可能是一种侵入性较小的方法，可以将治疗水平的溶酶体酶输送到大脑中。本项目的目的是确定野生型和改良型GUSB在小鼠和犬MPS VII模型中的分布以及组织病理学纠正的程度。我们还将确定溶酶体酶从眼睛到大脑的运输与储存减少是否与另一种溶酶体储存病相同。我们将通过以下具体目标来实现这些目标：1)我们将确定天然和修饰的GUSB的分布，以及玻璃体内注射基因转移载体后小鼠MPS VII模型CNS中溶酶体储存减少的程度。2)我们将测定天然GUSB和修饰GUSB的分布，以及玻璃体内注射基因转移载体后犬MPS VII模型CNS中溶酶体储存量减少的程度。3)我们将通过玻璃体内注射一种基因转移载体，测定棕榈酰蛋白硫酯酶1在幼年神经性蜡样脂褐膜病小鼠模型脑内的分布及对溶酶体储存的影响。

DESCRIPTION (provided by applicant): Cognitive deficits due to the accumulation of storage material in the brain can be the most severe clinical signs associated with lysosomal storage diseases. The CNS defects are also difficult to treat. We recently injected an AAV vector directly into the vitreous of the eye in the murine model of mucopolysaccharidosis type VII (MPS VII). High levels of GUSB activity were observed in the retina and in the optic nerve. We also discovered GUSB activity in the brains of NTS VII mice. GUSB activity was detected primarily in the visual tracts, specifically the lateral geniculate and superior colliculus. Interestingly, the reduction of lysosomal storage material extended beyond the visual system and into the hippocampus and cerebral cortex. Therefore, this approach may represent a less invasive method of delivering therapeutic levels of a lysosomal enzyme into the brain. The goals of this project are to determine the distribution of wild type and modified GUSB and the extent of histopathologic correction in both the mouse and canine models of MPS VII. We will also determine if the transport of a lysosomal enzyme from the eye to the brain with a reduction of storage is common to another lysosomal storage disease. We will accomplish these goals with the following Specific Aims. 1) We will determine the distribution of both native and modified GUSB, and the extent of lysosomal storage reduction in the CNS of the murine model of MPS VII following intravitreal injection of a gene transfer vector. 2) We will determine the distribution of both native and modified GUSB, and the extent of lysosomal Storage reduction in the CNS of the canine model of MPS VII following intravitreal injection of a gene transfer vector. 3) We will determine the distribution of palmitoyl protein thioesterase 1 and the effect on lysosomal storage in the brain of the murine model of infantile neuronal ceroid lipofuscinosis following intravitreal injection of a gene transfer vector.