Gene Therapy of the Gangliosidoses

描述（由申请方提供）：本项目的总体目标是在充分表征的真实猫人GM 1神经节苷脂沉积症模型中，使用最先进的慢病毒和腺相关病毒载体评价神经节苷脂沉积症病毒载体基因治疗的有效性和安全性。致死性神经病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系统成分。因此，我们将评估在子宫内和出生后非常早期给予病毒载体后中枢神经系统（CNS）和非CNS器官的转染。将表达标记或治疗基因的载体注射到妊娠25-50天的胎儿小猫或2-6周龄的围产期小猫的脑中或全身。将评价正常小猫的载体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GM 1神经节苷脂沉积症的小猫将在子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，并使用各种形态学、生物化学和分子方法评价神经系统和全身性疾病的进展。肝脏和胸腺疾病状态的评估将提供全身和CNS反应之间的比较。将在体外和体内研究基因治疗可能通过直接转染酶缺陷细胞或通过酶供体细胞交叉校正来改变神经节苷脂沉积过程的机制。将仔细监测病毒载体和转基因的不良反应。这项工作的结果将提供对溶酶体疾病和其他神经系统全局退行性疾病的基因治疗的潜在益处和风险的基本理解。这项临床前研究将产生必要的信息，导致基因治疗的翻译与神经元病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.