CORRECTION OF INHERITED PROTEIN DEFICIENCEIS BY GENE THERAPY

我们正在描述与影响神经系统的疾病有关的蛋白质，以便分离cDNA和基因组DNA，这些DNA可用于开发利用基因转移治疗疾病的方法。特别适合于基因治疗的初始尝试是那些疾病的全身性，在某些情况下甚至是神经系统的表现可以用重组改变的、可获得的骨髓、成纤维细胞、肝细胞或内皮细胞来治疗。我们已经使用了溶酶体紊乱戈谢病作为原型来开发更有效的基因转移，特别是使用小鼠模型。最近，我们用胚胎干细胞同源重组的方法建立了一个寿命长、病情轻的戈谢病小鼠模型。逆转录病毒载体已被用于在小鼠和患者细胞系和组织中表达人葡萄糖脑苷酶和其他基因，并逆转戈谢病小鼠模型组织中的脂质储存。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转移到特定组织是正在研究的策略之一。在子宫内治疗小鼠可能需要有效治疗影响神经系统的疾病，其中组织损伤发生在妊娠早期。这项研究的最初目标是将基因治疗应用于在某些情况下也会影响神经系统的非神经病变表型的疾病。我们还使用逆转录病毒介导的神经递质合成酶的转移，如酪氨酸羟化酶，用于体外和体内多巴缺乏状态的纠正。产生酪氨酸羟化酶的重组工程细胞（如成纤维细胞）已被用作左旋多巴释放的仓库，并已被移植到动物模型的神经系统中。当我们对遗传性神经和精神疾病的发病机制的理解提高，以及基因在特定组织和细胞中的转移和表达技术变得更加可预测时，我们应该能够将基因疗法的使用扩展到治疗更多影响神经系统的疾病。-基因治疗，戈谢病，小鼠，同源重组，胚胎干细胞，逆转录病毒载体，AAV，慢病毒，寡核苷酸

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. - gene therapy, Gaucher disease, mice, homologous recombination, ES cells, retroviral vectors, AAV, lentivirus, oligonucleotide