GROWTH FACTOR ENHANCED RETROVIRAL GENE TRANSFER TO CNS

描述：(改编自申请人的摘要)而逆转录病毒载体 可能会被整合为一种传递遗传物质的工具 进入中枢神经系统后，这些载体在大脑和脊髓中的使用受到限制 脐带由于缺乏旺盛的细胞分裂能力。申请人推测 基于MLV的病毒载体的这种固有限制可能是 通过利用几种不同生长的已知促有丝分裂作用而克服的 影响中枢神经系统细胞的因素。不同生长因子对细胞分裂的影响 不同的细胞类型。利用这一信息，PI假设一个人在 体内应用生长因子可促进基于MLV的基因转移 成人的大脑。本申请的目标是进一步研究这一点 将基因转移到大脑的技术方法，这是一个数量很少的区域 即使在受伤或疾病状态下也能分裂细胞。在目标一号中， 申请人建议确定生长因子的剂量反应曲线 (如：成纤维细胞生长因子、PDGF、EGF)诱导有丝分裂。这些研究将有助于建立 体内优化细胞分裂所需的生长因子浓度。目标 2将鉴定和量化在动物体内转导的特定细胞类型。 先注射一种特定的生长因子，然后注射 以MLV为基础的载体表达标记基因。这些实验将允许 申请人须决定给予生长因子是否能令人 针对特定类型的细胞将基因输送到中枢神经系统。《目标3》中的动物 用生长因子和MLV载体处理后，将允许存活长达 六个月。在这些动物体内转导的细胞类型将被确定并 量化，以便能够评估每个转基因的寿命 可以作为靶点的细胞群体。Aim 4将测试治疗方法 基因转移法在帕金森病大鼠模型中的应用 疾病。在这个模型中，生化和行为变化与 我们将对这种治疗进行分析。希望这些拟议的实验 将导致新一代研究，旨在扩大 MLV载体包括其他中枢神经系统退行性疾病模型。

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.