COMPREHENSIVE GENE THERAPY FOR MUCOPOLYSACHHARIDOSIS VII

粘多糖症综合基因治疗 VII

• 批准号: 7058335
• 负责人: JAYANTA ROY-CHOWDHURY
• 金额: $ 32.15万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7058335
• 关键词: beta glucuronidase; electron microscopy; enzyme deficiency; gene expression; gene therapy; laboratory mouse; mucopolysaccharides; mucopolysaccharidosis; polymerase chain reaction; therapy design /development

DESCRIPTION (provided by applicant): Our long-term objective is to develop safe and effective methods for comprehensive gene-based therapies for mucopolysaccharidoses, which are characterized by lysosomal glycosaminoglycan (GAG) deposition, resulting from inherited deficiency of enzymes needed for stepwise degradation of GAGs. Because GAG deposits affect multiple systems, including the liver, spleen, bones, brain and eyes, gene therapy for these disorders present a special challenge. In this project, we will target mucopolysaccharidosis-VTI (MPS-VII, beta-glucuronidase deficiency, Sly syndrome), since a well-characterized murine model is available. In preliminary studies, we have shown that transfer of the beta-glucuronidase (GUSB) gene using recombinant SV40-based vectors has the potential to ameliorate the GAG deposits in the brain as well as in the liver and spleen. The first specific aim is to optimize SV40- based vectors for safe and efficient systemic gene transfer. Novel viral gene-deleted vectors will be generated to accommodate promoters for efficient long-term transgene expression in various tissues. Insulators will be introduced to protect neighboring genes in the host genome from being accidentally activated by enhancers within the vector. In Specific Aim 2, we will determine optimal routes of administration, efficacy of gene transfer, tissue and serum GUSB levels and reduction of GAG-containing lysosomal vesicles. In addition to evaluating phenotypic correction of the liver and spleen, special emphasis will be placed on gene transfer to various types of brain cells in different regions of the brain after intravenous, intracerebro-ventricular or intracerebral injection of the vectors. We will also determine the potential of injecting these vectors into newborn or fetal mice on the ability to correct GUSB deficiency early in development, thereby preventing early brain damage, which is common in MPS VII. Since SV-40-based vectors are non-toxic, non-immunogenic and capable of integrating into the host genome, successful completion of this project should provide a safe and effective method for comprehensive molecular therapy of all forms of lysosomal storage disorders that are associated with visceral, skeletal and central nervous system injury.

描述（由申请人提供）：我们的长期目标是开发安全有效的方法，用于针对粘多糖病的综合基因治疗，其特征是溶酶体糖胺聚糖（GAG）沉积，这是由于逐步降解糖胺聚糖所需的酶的遗传性缺陷所致。 GAG。由于 GAG 沉积物影响多个系统，包括肝脏、脾脏、骨骼、大脑和眼睛，因此这些疾病的基因治疗面临着特殊的挑战。在这个项目中，我们将针对粘多糖贮积症-VTI（MPS-VII、β-葡萄糖醛酸酶缺乏症、Sly 综合征），因为已有一个特征良好的小鼠模型。在初步研究中，我们已经表明，使用基于 SV40 的重组载体转移 β-葡萄糖醛酸酶 (GUSB) 基因有可能改善大脑以及肝脏和脾脏中的 GAG 沉积。第一个具体目标是优化基于 SV40 的载体，以实现安全有效的系统基因转移。将产生新型病毒基因缺失载体，以容纳启动子，以便在各种组织中有效地长期表达转基因。将引入绝缘子以保护宿主基因组中的邻近基因免于被载体内的增强子意外激活。在具体目标 2 中，我们将确定最佳给药途径、基因转移功效、组织和血清 GUSB 水平以及含有 GAG 的溶酶体囊泡的减少。除了评估肝脏和脾脏的表型校正外，还将特别强调在静脉内、脑室内或脑内注射载体后，基因转移到大脑不同区域的各种类型的脑细胞。我们还将确定将这些载体注射到新生或胎儿小鼠体内的潜力，以纠正发育早期的 GUSB 缺陷，从而防止早期脑损伤，这在 MPS VII 中很常见。由于基于SV-40的载体无毒、无免疫原性并且能够整合到宿主基因组中，因此该项目的成功完成将为所有形式的溶酶体贮积症相关的综合分子治疗提供一种安全有效的方法。伴有内脏、骨骼和中枢神经系统损伤。