Gene Therapy of the Gangliosidoses

描述(由申请人提供)：该项目的主要目标是评估病毒载体基因治疗神经节苷脂增多症的有效性和安全性，使用最先进的慢病毒和腺相关病毒载体在具有良好特征的、真实的人类神经节苷脂增多症的猫科动物模型中应用。致命的神经病理性溶酶体疾病必须尽早治疗，以预防或逆转这些疾病的神经部分。因此，我们将评估在宫内和出生后极早期注射病毒载体后中枢神经系统(CNS)和非中枢神经系统器官的转染率。表达标记或治疗基因的载体将被注射到怀孕25-50天的胎猫或2-6周大的围产期猫咪的脑内或系统中。将评估正常小猫的矢量化标记基因的表达和分布，以提供用于优化治疗的基本生物学特征。患有GM1神经节苷脂增多症的小猫将在宫内和出生后使用表达溶酶体β-半乳糖苷酶的载体进行治疗，并使用各种形态、生化和分子方法评估神经和系统疾病的进展。评估肝脏和胸腺的疾病状态将提供全身反应和中枢神经系统反应之间的比较。基因治疗可能通过直接转染酶缺陷细胞或酶供体细胞交叉纠正来改变神经节苷脂病的进程的机制将在体外和体内进行研究。对病毒载体和转基因的不良反应将进行仔细监测。这项工作的结果将使人们对溶酶体疾病和其他全球神经系统退行性疾病的基因治疗的潜在好处和风险有基本的了解。这项临床前研究将产生必要的信息，从而将基因治疗转化为患有神经病理性溶酶体疾病的儿童。

DESCRIPTION (provided by applicant): The overarching goal of this project is to evaluate the application of virally vectored gene therapy of the gangliosidoses for efficacy and safety using state of the art lentiviral and adeno-associated viral vectors in a well characterized, authentic feline model of human GM1 gangliosidoses. Fatal neuronopathic lysosomal diseases must be treated at the earliest time possible to prevent or reverse the neurological component of these disorders. Therefore, we will evaluate transfection of central nervous system (CNS) and non-CNS organs after administration of viral vectors in utero and very early postnatally. Vectors expressing marker or therapeutic genes will be injected in the brain or systemically in fetal kittens at 25-50 days of gestation or perinatal kittens 2-6 weeks old. Normal kittens will be evaluated for expression and distribution of vectored marker genes to provide the essential biological characteristics used to optimize therapy. Kittens with GM1 gangliosidoses will be treated in utero and postnatally with vectors expressing lysosomal beta-galactosidase and evaluated for progression of neurological and systemic disease using a variety of morphological, biochemical and molecular methods. Assessment of disease status in liver and thymus will provide comparison between systemic and CNS responses. Mechanisms by which gene therapy may alter the course of the gangliosidoses by direct transfection of enzyme deficient cells or cross correction by enzyme donor cells, will be studied in vitro and in vivo. Adverse reactions to viral vectors and transgenes will be monitored carefully. The results of this work will provide basic understanding of the potential benefits and risks resulting from gene therapy of lysosomal diseases and other global degenerative diseases of the nervous system. This preclinical study will generate essential information leading to the translation of gene therapy to children with neuronopathic lysosomal diseases.