GROWTH FACTOR ENHANCED RETROVIRAL GENE TRANSFER TO CNS

描述：（改编自申请人的摘要）虽然逆转录病毒载体 可以作为一种传递遗传物质的工具 虽然这些载体在中枢神经系统中的应用受到限制， 因为缺乏强大的细胞分裂。申请人推测， 基于MLV的病毒载体的这种固有局限性可能潜在地 通过利用已知的几种不同生长的促有丝分裂效应来克服 对CNS细胞的影响。不同的生长因子影响细胞分裂， 不同的细胞类型利用这些信息，PI假设， 生长因子的体内应用可以增强基于MLV的基因转移到 成人大脑本申请的目标是进一步研究这一点。 基因转移到大脑的技术方法，大脑是一个数量很少的区域。 即使在受伤后或疾病状态期间也能分裂细胞。目标一， 申请人建议确定生长因子的剂量反应曲线 (eg., FGF、PDGF、EGF诱导有丝分裂。这些研究将有助于建立 这是优化体内细胞分裂所需的生长因子浓度。目的 2将鉴定和定量动物中转导的特定细胞类型 首先用特定的生长因子引发， 基于MLV的表达标记基因的载体。这些实验将使 申请人确定给予生长因子是否能够使人 靶向特定的细胞类型，用于将基因递送至CNS。目标3：动物 将允许用生长因子和MLV载体处理的小鼠存活长达 6个月将确定这些动物中转导的细胞类型， 定量以允许评估每个转基因的寿命。 可以被靶向的细胞群。目标4将测试治疗 基因转移方法在帕金森病大鼠模型中的潜力 疾病在这个模型中，与之相关的生化和行为变化 将对该处理进行分析。希望这些提议的实验 将导致新一代的研究，旨在扩大应用 MLV载体包括CNS的其他神经退行性疾病模型。

DESCRIPTION: (Adapted from the Applicant's Abstract) While retroviral vectors could potentially be incorporated as a tool for delivering genetic material into the CNS, the use of these vectors has been limited in the brain and spinal cord because of the lack of robust cell division. The applicant speculated that this inherent limitation of MLV based viral vectors could potentially be overcome by utilizing the known mitogenic effect of several different growth factors on cells of the CNS. Different growth factors effect cell division in different cell types. Using this information the PI has hypothesized that an in vivo application of growth factor could enhance MLV based gene transfer to the adult brain. The goal of the present application is to examine further this technical approach of gene transfer to the brain, a region with poor numbers of dividing cells even after injury or during disease states. In aim one, the applicant proposes to determine the dose response curves for growth factor (eg., FGF,PDGF, EGF) induced mitogenesis. These studies will help establish the concentration of growth factor required to optimize cell division in vivo. Aim 2 will identify and quantify the particular cell types transduced in animals first primed with a specific growth factor and subsequently given an injection of MLV based vectors expressing a marker gene. These experiments will allow the applicant to determine if the administration of a growth factor enables one to target specific cell types for gene delivery to the CNS. In aim 3, animals treated with growth factor and MLV vectors will be allowed to survive for up to 6 months. The cell types transduced in these animals will be determined and quantified so as to allow the assessment of longevity of the transgene for each of the cell populations that can be targeted. Aim 4 will test the therapeutic potential of the gene transfer approach in the rat model of Parkinson's disease. In this model the biochemical and behavioral changes associated with this treatment will be analyzed. It is hoped that these proposed experiments will lead to a new generation of studies designed to broaden the application of MLV vectors to include other neurodegenerative disease models of the CNS.