Glucocerebrosidase Gene Transfer to the Nervous System

描述（由申请人提供）：戈谢病的神经病变类型仍然对中枢神经系统症状的治疗提出挑战。一种方法是基因治疗，但是，尽管葡萄糖脑苷酶（GC） cDNA已经成功地通过逆转录病毒载体转移到正常小鼠和人类造血干细胞中，或通过腺相关病毒载体转移到肌肉细胞中，并且转导的小鼠细胞成功地移植到小鼠体内，迄今为止进行的三项临床试验结果要么是低水平的基因转移，要么是输注后植入失败，只有一个例外。利用逆转录病毒、慢病毒和其他载体，已经成功地证明了基因向神经元的转移。在受体动物的大脑中已经证实了转导的神经元和造血祖细胞的移植。我们假设移植造血或神经祖细胞由含有人糖脑苷酶cDNA的逆转录病毒和慢病毒载体转导。本研究将验证含有人葡萄糖脑苷酶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移植在受体Gaucher L444P突变小鼠体内并导致酶水平提高的假设。用逆转录病毒和慢病毒载体成功地证明了基因向神经元的转移，并且在受体动物的大脑中实现了转导的神经元和造血祖细胞的移植。然而，尽管在动物模型上取得了成功，但迄今为止进行的三项临床试验通常要么导致低水平的基因转移，要么导致细胞输注后植入失败。为此，我们将：1)更完整地描述新建立的Gaucher L444P突变小鼠；2)静脉注射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酶水平的改善程度；3)通过鞘内给药移植体外转导的造血或神经元祖细胞，以确定这些细胞是否会在大脑中重新填充并表达葡萄糖脑苷酶活性。在这项研究完成后，我们将确定慢病毒介导的人类GC基因转移是否会导致受体Gaucher突变小鼠组织（特别是大脑）中酶水平的持续改善。

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.