Glucocerebrosidase Gene Transfer to the Nervous System

描述（由申请方提供）：神经元病型戈谢病仍然是CNS症状治疗的挑战。一种方法是基因治疗，但是尽管事实上葡萄糖脑苷脂酶（GC）cDNA已经通过逆转录病毒载体成功地转移到正常的鼠和人造血干细胞或通过腺相关病毒载体转移到肌肉细胞，并且转导的鼠细胞成功地移植到小鼠中，但迄今为止进行的三项临床试验导致低水平的基因转移，或输注后植入失败，只有一个例外。使用逆转录病毒、慢病毒和其他载体已经成功地证明了向神经元的基因转移。已证明转导的神经元和造血祖细胞的植入在受体动物的脑中。我们假设，移植造血或神经元祖细胞转导的逆转录病毒和慢病毒载体含有人葡萄糖脑苷脂酶的cDNA。本研究将检验以下假设：移植由含有人葡萄糖脑苷脂酶cDNA的逆转录病毒和慢病毒载体转导的造血或神经元祖细胞将植入受体Gaucher L444 P突变小鼠中并导致酶水平的改善。已经成功地证明了逆转录病毒和慢病毒载体的基因转移到神经元，并且已经在受体动物的脑中实现了转导的神经元和造血祖细胞的植入。然而，尽管在动物模型中取得了成功，但迄今为止进行的三项临床试验通常导致基因转移水平低或细胞输注后植入失败。为了实现这一点，我们将：1）更完整地表征新建立的Gaucher L444 P突变小鼠; 2）静脉内施用逆转录病毒和慢病毒转导的造血祖细胞以确定受体突变小鼠组织中葡萄糖脑苷脂酶水平改善的程度;并且，在本发明中，第三章通过鞘内施用离体转导的造血祖细胞或神经元祖细胞进行移植，以确定这些细胞是否将在脑中重新增殖并表达葡糖脑苷脂酶活性。在本研究完成时，我们将确定慢病毒介导的人GC基因转移是否会导致受体高雪氏突变小鼠组织（特别是脑）中酶水平的持续和改善。

DESCRIPTION (provided by applicant): The neuronopathic types of Gaucher disease still present a challenge for the treatment of the CNS symptoms. One approach would be gene therapy, but despite the fact that the glucocerebrosidase (GC) cDNA has been successfully transferred by retroviral vector to normal murine and human hematopoietic stem cells or by adeno-associated virus vector to muscle cells, and the transduced murine cells successfully transplanted in mice, the three clinical trials conducted to date resulted in either low levels of gene transfer, or a failure of engraftment after infusion, with a single exception. Gene transfer to neurons has been successfully demonstrated using retroviral, lentiviral, and other vectors. Engraftment of transduced neuronal and hematopoietic progenitors has been demonstrated in the brain of recipient animals. We hypothesize that transplantation of hematopoietic or neuronal progenitor cells transduced by retroviral and lentiviral vectors containing the cDNA for human glucocerebrosidase. This study will test the hypothesis that transplantation of hematopoietic or neuronal progenitor cells transduced by retroviral and lentiviral vectors containing the cDNA for human glucocerebrosidase will engraft in recipient Gaucher L444P mutant mice and lead to improved enzyme levels. Gene transfer to neurons has been successfully demonstrated with both retroviral and lentiviral vectors, and engraftment of transduced neuronal and hematopoietic progenitors has been attained in the brains of recipient animals. However, despite this success in animal models, the three clinical trials conducted to date have generally resulted in either low levels of gene transfer or a failure of engraftment after cell infusion. To accomplish this, we will: 1) characterize more completely the newly established Gaucher L444P mutant mouse; 2) intravenously administer retrovirally and lentivirally transduced hematopoietic progenitors to determine the extent of improved glucocerebrosidase enzyme levels in the tissues of the recipient mutant mice; and, 3) transplant by intrathecal administration ex vivo transduced hematopoietic or neuronal progenitors to determine if these cells will repopulate in brain and express glucocerebrosidase activity. At the completion of this study we will have determined whether lentiviral mediated gene transfer of human GC will result in sustained and improved enzyme levels in tissues, particularly brain, of recipient Gaucher mutant mice.