GENE THERAPHY FOR PARKINSON'S DISEASE

帕金森病的基因治疗

• 批准号: 6133080
• 负责人: Martha D Bohn
• 金额: $ 31.7万
• 依托单位: LURIE CHILDREN'S HOSPITAL OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6133080
• 关键词: Parkinson's disease; disease /disorder model; dopamine receptor; gene expression; gene therapy; laboratory rat; neurons; neurotrophic factors; technology /technique development; tissue /cell culture; transfection /expression vector

DESCRIPTION: (Verbatim from the Applicant's Abstract) The long-term goal of this project is to develop novel gene therapies for neurodegenerative diseases. In the previous support period, we focused on adenoviral (Ad) vectors to deliver the gene encoding GDNF (glial cell line-derived neurotrophic factor). Ad-GDNF injected into either the substantia nigra or striatum of a progressive degeneration model of Parkinson's disease protected dopaminergic (DA) neurons against cell death induced by the neurotoxin 6-OHDA. Ad-GDNF injected into the striatum also prevented the acquisition of behaviors and molecular changes that occurred in DA deficient young and aged rats. This proposal focuses on the hypothesis that anti-apoptotic gene delivery will also protect DA neurons in vitro and in vivo and have a synergistic effect with delivery of neurotrophic factor genes. Viral vectors harboring genes that block specific apoptotic death pathways, including XIAP, a dominant-negative caspase-9, bcl-2 and bclxl will be studied for effects on survival and function of DA neurons either alone or in combination with neurotrophic factors, GDNF or neurturin. Genes will be delivered to DA neurons in culture and in rat brain using helper free HSV:AAV hybrid amplicon vectors. These vectors will incorporate bidirectional expression cassettes that drive both the therapeutic gene and the cellular marker gene, green fluorescent protein, to permit specific evaluation of transduced cells. Expression will be controlled using the tetracycline responsive element such that transgene expression is "on" in the presence of tetracycline activator (TA) and in the absence of doxcycline (Dox). Vectors will be made in which TA is driven by a viral promoter of the DA cellular promoter, tyrosine hydroxylase (TH). Effects of the 'therapeutic' genes will be studied using non-neuronal cells, the DA cell line, MN9D, and primary fetal DA neurons treated with the neurotoxins, MPP+ or 6-OHDA or other cellular insults. In vivo effects of therapeutic genes will be studied in: 1) rats that have received grafts of fetal DA neurons, and 2) rats that have received a progressive 6-OHDA lesion of the nigrostriatal projection. Reversibility of effects will be studied by administration of Dox. Effects on DA neurons will be evaluated using quantitative morphometric and molecular techniques and behavioral evaluations. This project also aims to continue its evaluation of new generation viral vectors, including E2b deleted Ad, totally gutted Ad, and HSV:AAV amplicon, for stability and levels of expression in the nigrostriatal system. The studies involve collaborations among investigators at Children's Memorial Hospital and Northwestern Univ. Med. School and are relevant to the development of novel therapies for neurodegenerative diseases and injuries to the CNS.

描述：(逐字摘自申请人的摘要)的长期目标 该项目旨在开发治疗神经退行性疾病的新型基因疗法。 在之前的支持期间，我们专注于腺病毒(Ad)载体，以 携带胶质细胞源性神经营养因子(GDNF)编码基因。 进展性黑质或纹状体内注射Ad-GDNF 帕金森病保护的多巴胺能神经元变性模型 抗神经毒素6-OHDA诱导的细胞死亡。Ad-GDNF注射入脑内 纹状体也阻止了行为和分子变化的获得 发生于DA缺乏的幼年和老年大鼠。这项提案的重点是 抗凋亡基因传递也将保护多巴胺能神经元的假说 并与神经营养递送有协同作用 因子基因。携带阻断特定细胞凋亡的基因的病毒载体 包括XIAP、显性负性caspase-9、bcl2和bclx1在内的通路将 单独或联合研究对多巴胺神经元存活和功能的影响 联合应用神经营养因子、神经营养因子或神经节苷脂。基因将会是 用无辅助子HSV：AAV向培养和大鼠脑内的DA神经元传递 杂交扩增子载体。这些向量将包含双向 同时驱动治疗性基因和细胞的表达盒 标记基因，绿色荧光蛋白，允许特异性评估 转导细胞。表达将使用四环素进行控制 反应元件，使转基因表达在存在的情况下是“开启”的 四环素激活剂(TA)和在没有多西环素(Dox)的情况下。向量 其中TA是由DA细胞的病毒启动子驱动的 酪氨酸羟化酶(TH)启动子。“治疗性”基因的作用将是 使用非神经细胞、DA细胞系、MN9D和原代胎儿DA进行研究 用神经毒素、MPP+或6-OHDA或其他细胞损伤处理神经元。 治疗性基因的体内效应将在以下方面进行研究：1)具有 接受胚胎DA神经元移植，以及2)接受了 进行性黑质纹状体投射的6-羟色胺损伤。的可逆性 影响将通过Dox的管理进行研究。对DA神经元的影响将是 使用定量形态测量和分子技术进行评估 行为评估。该项目还旨在继续评价 新一代病毒载体，包括E2b删除的广告，完全清除的广告，以及 HSV：AAV扩增子，对黑质纹状体的稳定性和表达水平有影响 系统。这些研究涉及儿童医院的调查人员之间的合作 纪念医院和西北大学。地中海医院。学校，并与 神经退行性疾病和损伤的新疗法的开发 中枢神经系统。