Global spinal cord trophic factor delivery using intracerebral rAAV in ALS rats

描述（由申请人提供）：肌萎缩性侧索硬化症（ALS）是一种进行性和致死性神经系统疾病，其特征是上下运动神经元死亡。没有已知的治疗方法。我们的团队有使用病毒载体在神经系统疾病动物模型中递送蛋白质的经验，并已在其他医学适应症中进行临床试验。最近的报道表明，胶质细胞系源性神经营养因子（GDNF）和胰岛素样生长因子1（IGF1）是可能有效治疗ALS的营养因子。然而，由于ALS是进行性的，将需要慢性蛋白质递送。而且，受累的运动神经元分布在整个脊髓的前角。慢性递送和全局脊髓递送是蛋白质递送的主要挑战。该项目的目标是使用重组腺相关病毒载体，注射到单个脑区，通过已知的解剖学脊髓投射束利用顺行运输将分泌的营养因子GDNF和IGF 1递送到整个脊髓。本发明的总体假设是，运动皮质的锥体细胞或红核中的神经元可以被重组腺相关病毒载体（rAAV）永久转导以产生GDNF和IGF 1，并且这些蛋白质可以以治疗方式递送至整个脊髓。因此，本研究的目的有两个：1）观察rAAV 1、rAAV 5和rAAV 8注入正常大鼠红核后，报告基因、绿色荧光蛋白（GFP）、IGF 1和GDNF顺行转运的效率、持续时间和安全性。以优化基因产物的脊髓传递。2)在肌萎缩侧索硬化大鼠模型中，研究使用rAAV递送的GDNF和IGF 1拯救下运动神经元疾病的潜力。将通过评价对照和ALS大鼠中运动症状的进展以及通过脊髓的详细组织病理学来确定挽救。目前的建议可能会产生大量的一般信息载体介导的蛋白质传递到整个脊髓。这些数据可能对脊髓损伤，特别是ALS具有普遍意义。

DESCRIPTION (provided by applicant): Amyotrophic Lateral Sclerosis (ALS) is a progressive and lethal neurological disorder characterized by death of upper and lower motoneurons. There is no known cure. Our group has experience using viral vectors to deliver proteins in animal models of neurological disorders and have proceeded to clinical trials in other medical indications. Recent reports indicate that glial cell line- derived neurotrophic factor (GDNF) and insulin-like growth factor 1 (IGF1) are trophic factors that may be efficacious in treating ALS. However, since ALS is progressive, chronic protein delivery will be needed. Moreover, the affected motoneurons are distributed in the ventral horn of the entire spinal cord. Chronic delivery and global spinal cord delivery are major challenges for protein delivery. The goal of this project is to use recombinant adeno-associated viral vectors, injected in single brain regions, to deliver the secreted trophic factors GDNF and IGF1 to the entire spinal cord, by using anterograde transport via known anatomical spinal projecting tracts. The overall hypothesis of the present proposal is that either the pyramidal cells of the motor cortex or the neurons in the red nucleus can be permanently transduced by recombinant adeno-associated viral vectors (rAAV) to produce GDNF and IGF1 and these proteins can be delivered to the entire spinal cord in a therapeutic manner. Therefore, two aims are proposed: 1) To determine the efficiency, duration and safety of anterograde transport of the reporter gene, green fluorescent protein (GFP), IGF1, and GDNF after injection of rAAV1, rAAV5 and rAAV8 into the red nucleus in normal rats. In order to optimize spinal cord delivery of gene products. 2) To examine the potential for rescue of lower motor neuron disease in an amyotrophic lateral sclerosis rat model using rAAV-delivered GDNF and IGF1. Rescue will be determined both by evaluating the progression of the motor symptoms in control and ALS rats and by detailed histopathology of the spinal cord. The present proposal is likely to produce a significant amount of general information about vector-mediated protein delivery to the entire spinal cord. These data may have general implications for spinal cord injuries and ALS in particular.