Gene therapy for ataxia

共济失调的基因治疗

• 批准号: 6861183
• 负责人: R. Scott McIvor
• 金额: $ 17.91万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6861183
• 关键词: RNA interference; adeno associated virus group; antisense nucleic acid; cerebellar Purkinje cell; cerebellar ataxia /dyskinesia; fibroblast growth factor; gene therapy; genetically modified animals; growth factor receptors; laboratory mouse; nerve /myelin protein; nonhuman therapy evaluation; nuclear proteins; pathologic process; proteoglycan; ribozymes; tissue /cell culture; transfection; transfection /expression vector

The cerebellum is a region of the brain that plays a major role in the control of movement and cognition. Spinocerebellar Ataxia Type 1 (SCA1) is a fatal human disorder that is one of at least five inherited ataxias, and a total of at least eight neurodegenerative diseases, each caused by the expression of a mutant protein containing an expanded polyglutamine region. Pathogenesis is due to a gain of function conferred by mutant ataxin-1 protein and has been recapitulated in a transgenic mouse model resulting in degeneration of neuronal Purkinje cells in the cerebellum. As a potential approach toward gene therapy for SCA1, we demonstrated during the current period of study: (i) That adeno-associated virus vectors (AAV) are an effective tool for in vivo gene transfer into cerebellar Purkinje cells, yielding up to 3% transduction in a cerebellar hemisphere after a single intracerebellar injection; (ii) That ribozymes and antisense RNAs targeting specific positions in the ataxin message destabilize ataxin message in vitro and in tissue culture cells in vivo, respectively. In this competing renewal application, we propose to extend these studies through the execution of 4 specific aims: (i) Further characterization and optimization of AAV-mediated Purkinje cell transduction in vivo, including the molecular role of basic fibroblast growth factor receptor 1 and heparin sulfate proteoglycan in virus binding, and the relative effectiveness of AAV serotypes 1, 2 and 5 in mediating transduction of cerebellar Purkinje cells; (ii) Further testing of molecular approaches for downregulating or correcting expanded ataxin-1 message, including antisense, ribozyme, trans-splicing and RNA interference; (iii) Establishment of anti-ataxin transgenes, subsequently crossing with SCA1 transgenic mice to evaluate the effectiveness of these approaches in controlling Purkinje cell degeneration and development of ataxia; and (iv) Use of AAV vectors to deliver anti-ataxin or ataxin correcting sequences to cerebellar Purkinje cells in SCA1 transgenic mice, as a direct model of gene therapy for SCA1 in humans. Results from these studies will thus provide optimized molecular tools and delivery conditions which will be applicable to treatment not only of SCA1 but other dominant-acting and other neurologic disorders as well.

小脑是大脑的一个区域，在控制运动和认知方面发挥着重要作用。脊髓小脑性共济失调 1 型 (SCA1) 是一种致命的人类疾病，是至少五种遗传性共济失调和总共至少八种神经退行性疾病之一，每种疾病都是由含有扩展的多聚谷氨酰胺区域的突变蛋白的表达引起的。发病机制是由于突变的ataxin-1蛋白赋予的功能获得，并在转基因小鼠模型中得到了重现，导致小脑神经元浦肯野细胞变性。 作为 SCA1 基因治疗的潜在方法，我们在当前的研究期间证明：(i) 腺相关病毒载体 (AAV) 是体内基因转移至小脑浦肯野细胞的有效工具，产量高达 3%单次小脑内注射后小脑半球的转导； (ii)靶向ataxin信息中特定位置的核酶和反义RNA分别在体外和体内组织培养细胞中破坏atataxin信息的稳定性。在这个竞争性的更新应用中，我们建议通过执行 4 个具体目标来扩展这些研究：（i）进一步表征和优化 AAV 介导的浦肯野细胞体内转导，包括碱性成纤维细胞生长因子受体 1 和肝素的分子作用病毒结合中的硫酸盐蛋白多糖， 以及 AAV 血清型 1、2 和 5 在介导小脑浦肯野细胞转导中的相对有效性； (ii) 进一步测试下调或纠正扩展的ataxin-1信息的分子方法，包括反义、核酶、反式剪接和RNA干扰； (iii) 建立抗共济失调蛋白转基因，随后与 SCA1 转基因小鼠杂交，以评估这些方法在控制浦肯野细胞变性和共济失调发展方面的有效性； (iv)使用AAV载体将抗共济失调蛋白或共济失调蛋白校正序列递送至SCA1转基因小鼠的小脑浦肯野细胞，作为 人类 SCA1 基因治疗的直接模型。因此，这些研究的结果将提供优化的分子工具和递送条件，这些工具和递送条件不仅适用于 SCA1 的治疗，也适用于其他主导作用和其他神经系统疾病的治疗。