Gene Therapy of the Gangliosidoses

神经节苷脂病的基因治疗

• 批准号: 6643315
• 负责人: HENRY J. BAKER
• 金额: $ 18.13万
• 依托单位: AUBURN UNIVERSITY AT AUBURN
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-07 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6643315
• 关键词: Adenoviridae; Lentivirus; beta galactosidase; cats; central nervous system; complementary DNA; disease /disorder; embryo /fetus therapy; enzymes; gangliosidosis; gene therapy; green fluorescent proteins; lysosomes; neuropathology; newborn animals; nonhuman therapy evaluation; polymerase chain reaction; transfection /expression vector

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.

描述（由申请人提供）：该项目的总体目标是在特征明确、真实的人类 GM1 神经节苷脂猫科动物模型中，使用最先进的慢病毒和腺相关病毒载体，评估神经节苷脂病病毒载体基因治疗的有效性和安全性。致命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产后早期施用病毒载体后中枢神经系统 (CNS) 和非 CNS 器官的转染情况。表达标记或治疗基因的载体将被注射到妊娠 25-50 天的胎儿小猫或 2-6 周大的围产期小猫的大脑或全身。将评估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 GM1 神经节苷脂增多症的小猫将在子宫内和出生后接受表达溶酶体 β-半乳糖苷酶的载体治疗，并使用各种形态学、生化和分子方法评估神经和全身疾病的进展。对肝脏和胸腺疾病状态的评估将提供全身反应和中枢神经系统反应之间的比较。将在体外和体内研究基因治疗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病进程的机制。将仔细监测对病毒载体和转基因的不良反应。这项工作的结果将为溶酶体疾病和其他神经系统全球退行性疾病的基因治疗带来的潜在益处和风险提供基本了解。这项临床前研究将产生重要信息，从而将基因疗法转化为患有神经病性溶酶体疾病的儿童。