Gene delivery methods for global therapy in the CNS

描述（由申请人提供）：溶酶体贮积病（lsd）是最常见的儿童遗传疾病类型，在普通人群中估计频率为1 / 7700。据估计，60%的迷幻药都有一定程度的神经损伤，目前尚无治疗方法。因此，探索治疗神经性lsd的新方法是至关重要的。这项工作将重点关注gm1神经节苷脂病，这是一种无法治疗的儿童疾病，由溶酶体酸β -半乳糖苷酶（b-gal）缺乏引起的中度至重度神经功能障碍，伴有gm1神经节苷脂在中枢神经系统的积累。我们将在患有gm1神经节脂质沉积症的年轻成年小鼠中进行这些研究，因为大多数LSD病例是在儿童早期或后期被诊断出来的，那时大脑的发育与年轻成年或成年小鼠相当。我们实验室和其他实验室的实验表明，AAV8载体在将基因转移到成年小鼠大脑方面非常有效。此外，我们的研究表明，AAV8载体经尾静脉血管内输注后可以转导脑微血管内皮。因此，AAV8载体将在整个实验中使用。本研究旨在验证以下两个假设：1)成年GM1-神经节脂质病小鼠丘脑中小鼠b-gal的从头表达足以完全纠正整个CMS的病理生化异常，并在神经运动功能测试中表现正常；2) GM1神经节脂病小鼠脑微血管内皮表达b-半乳糖可导致整个中枢神经系统中GM1-和GA1-神经节脂苷的野生型水平，在神经运动功能测试中表现正常。本研究将编码b-gal或GFP的AAV8载体双侧注射到2月龄GM1-神经节脂质病小鼠的丘脑中（目的1），或在内皮或肝脏特异性启动子的控制下，通过尾静脉递送编码b-gal或GFP的AAV8载体（目的2）。治疗组和对照组小鼠将在1年内的不同时间点评估神经病理、生化、免疫和行为参数。我们预计这些实验的发现将适用于其他lsd模型，并且在不久的将来，它们可以在进入临床试验之前在gm1神经节脂质沉积症的大型动物模型中进行评估。

DESCRIPTION (provided by applicant): Lysosomal storage diseases (LSDs) comprise the most common type of childhood genetic disorder, with an estimated frequency of 1 in 7700 in the general population. It is estimated that 60% of all LSDs have some degree of neurological involvement for which no treatment is available. Thus it is of paramount importance to investigate new approaches to treat neuronopathic LSDs. This work will focus on GM1-gangliosidosis, which is an untreatable childhood disease with moderate to severe neurological impairment caused by a deficiency of lysosomal acid beta-galactosidase (b-gal) with accumulation of GM1-ganglioside in the central nervous system. We will conduct these studies in young adult mice with GM1-gangliosidosis because most LSD cases are diagnosed in early childhood or later when the brain is developmentally equivalent to young adult or adult mice. Experiments in our laboratory and others have shown that AAV8 vectors are exceptionally efficient for gene transfer to the adult mouse brain. Moreover, our studies have shown that AAV8 vectors can transduce the brain microcapillary endothelium after intravascular infusion via the tail vein. Thus AAV8 vectors will be used throughout these experiments. The aims in this application were designed to test the following two hypotheses: 1) De novo expression of mouse b-gal in the thalamus of adult GM1- gangliosidosis mice is sufficient to achieve complete correction of patho-biochemical abnormalities throughout the CMS and normal performance in tests of neuro-motor function; 2) Expression of b-gal in the brain microcapillary endothelium of GM1-gangliosidosis mice will result in wild type levels of GM1- and GA1- ganglioside throughout the CNS and normal performance in tests of neuro-motor function. Here we will inject AAV8 vectors encoding for b-gal or GFP bilaterally into the thalamus of 2 month old GM1- gangliosidosis mice (Aim 1) or deliver AAV8 vectors encoding b-gal or GFP under control of endothelial- or liver-specific promoters via the tail vein (Aim 2). Treated and control mice will be evaluated for neuropathological, biochemical, immunological, and behavioral parameters at different time points over 1 year. We anticipate the findings from these experiments to be applicable to other models of LSDs, and that in the near future they can be evaluated in a large animal model of GM1-gangliosidosis before moving to clinical trials.