Development of integrating HSV amplicons for Parkinson's disease

帕金森病整合 HSV 扩增子的开发

• 批准号: 6690914
• 负责人: WILLIAM J. BOWERS
• 金额: $ 16.36万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6690914
• 关键词: 6 hydroxydopamine; Alphaherpesvirinae; Parkinson's disease; biotechnology; cooperative study; corpus striatum; disease /disorder model; experimental brain lesion; gene delivery system; gene therapy; laboratory rat; neuroprotectants; neurotrophic factors; reporter genes; substantia nigra; transposon /insertion element

Gene transfer methodologies have created the opportunity for developing gene therapy for human neurological diseases such as Parkinson's Disease (PD). Adeno-associated virus (AAV) and lentivirus vectors, both of which integrate into the host cell chromosome, have been shown to provide long-term expression of genes and therapeutic efficacy in animal models of PD. These vectors, however, exhibit a number of disadvantages, among them an inability to harbor large transgene segments. Gene therapy vectors based upon the Herpes Simplex virus (HSV) offer numerous advantages for the development of novel therapeutics for PD. These include broad cellular tropism, large DNA packaging capacity that allows for expression of multiple genes, and high transduction efficiency. We propose to modify HSV-derived amplicon vectors to provide stable nigrostriatal cell-specific gene expression. For this pipeline project, we will initially construct novel integrating forms of the amplicon that will direct expression of a reporter gene product specifically within cells of the striatum and substantia nigra. DNA insulator elements will be introduced to facilitate maintenance of the integrated transcription unit in a euchromatic state. These new vectors will be co-administered with an amplicon expressing the Sleeping Beauty transposase into the striata of unlesioned rats, where gene expression duration and cell-type specificity will be experimentally derived. The vectors will then be used to deliver glial cell line-derived neurotrophic factor (GDNF) prior to 6-OHDA-induced lesioning of rats to assess the capacity of the new vector system to confer protection to the nigrostriatal pathway. The proposed studies will yield a novel HSV vector system, provide a detailed understanding of transgene expression in vivo, and evaluate its therapeutic effectiveness in protecting nigrostriatal neurons in a well-established rodent model of PD.

基因转移方法为开发人类神经系统疾病如帕金森病（PD）的基因治疗创造了机会。腺相关病毒（AAV）和慢病毒载体均可整合到宿主细胞染色体中，在PD动物模型中已显示出长期的基因表达和治疗效果。然而，这些载体表现出许多缺点，其中包括无法容纳大的转基因片段。基于单纯疱疹病毒（HSV）的基因治疗载体为帕金森病的新疗法的开发提供了许多优势。这些包括广泛的细胞趋向性，大的DNA包装能力，允许多个表达