CORRECTION OF INHERITED PROTEIN DEFICIENCEIS BY GENE THERAPY

我们正在研究与疾病有关的蛋白质 影响神经系统以允许cDNA的分离， 可用于开发治疗疾病的基因组DNA 使用基因转移。特别适合基因改造的初步尝试 治疗是那些全身性的疾病， 在某些情况下，即使是神经系统的表现， 可以用重组改变的，可接近的骨髓治疗， 成纤维细胞、肝或内皮衍生细胞。我们已经使用了 以溶酶体疾病戈谢病为原型发展起来的 有效的基因转移，特别是使用小鼠模型。最近 我们已经建立了一个长寿的，轻度感染的小鼠模型， 胚胎干同源重组引起的戈谢病 细胞逆转录病毒载体已用于表达人源性逆转录病毒。 小鼠和患者细胞系中的葡糖脑苷脂酶和其他基因 和组织，并逆转脂质在小鼠组织中的储存， 戈谢病的模型。此外，受体、脂质体、 寡核苷酸、腺相关病毒（AAV）和慢病毒 介导的DNA转移到特定组织中是策略之一， 正在调查可能需要在小鼠中进行子宫内治疗， 有效治疗影响神经系统的疾病， 组织损伤发生在妊娠早期。这个项目的最初目标是 研究是将基因治疗应用于非神经病患者 在某些情况下，这些疾病的表型也会影响 神经系统我们还使用了逆转录病毒介导的转移， 神经递质合成酶如酪氨酸羟化酶 对于多巴缺乏的体外和体内校正 states.生物工程细胞（例如，成纤维细胞） 生产酪氨酸羟化酶的人已经被用作 左旋多巴释放并已被移植到神经 动物模型系统。当我们理解 遗传性神经精神疾病的发病机制 疾病的改善，并作为技术转让， 基因在特定组织和细胞中的表达变得越来越多， 可以预见，我们应该能够将基因治疗的使用扩展到 治疗大量影响神经系统的疾病 系统

We are characterizing proteins involved in disorders affecting the nervous system to permit the isolation of cDNA and genomic DNA that can be used to develop treatments disorders using gene transfer. Particularly suited for initial attempts at gene therapy are those disorders where the systemic and in some instances even the neurologic manifestations of the disorder are treatable using recombinantly altered, accessible bone marrow, fibroblast, hepatic, or endothelial derived cells. We have used the lysosomal disorder Gaucher disease as a prototype to develop more efficient gene transfer, particularly using mouse models. Recently we have produced a long lived, mildly affected, mouse model of Gaucher disease by homologous recombination in embryonic stem cells. Retroviral vectors have been used to express human glucocerebrosidase and other genes in mouse and patient cell lines and tissues, and to reverse storage of lipid in tissues in murine models of Gaucher disease. In addition, receptor, liposome, oligonucleotide, adeno-associated virus (AAV) and lentivirus mediated DNA transfer into specific tissues are among strategies being investigated. In utero treatments in mice may be required for effective treatment of disorders affecting the nervous system where tissue damage occurs early in gestation. An initial goal of this research is the application of gene therapy to the non-neuronopathic phenotypes of disorders that can, in some cases, also affect the nervous system. We have also used retroviral mediated transfer of neurotransmitter synthesizing enzymes such as tyrosine hydroxylase for both the in-vitro and in-vivo correction of DOPA deficiency states. Recombinantly engineered cells (for instance, fibroblasts) producing tyrosine hydroxylase have been used as depots of L-DOPA release and have been transplanted into the nervous system of animal models. When our understanding of the pathogenetic mechanisms of inherited neurologic and psychiatric disease improves and as technologies for the transfer and expression of genes in specific tissues and cells becomes more predictable, we should be able to extend the use of gene therapy to treatment of a larger number of disorders affecting the nervous system.