Glucocerebrosidase Gene Transfer to the Nervous System

描述(由申请人提供)：神经病变类型的高谢病对中枢神经系统症状的治疗仍然是一个挑战。一种方法是基因治疗，但尽管已成功地通过逆转录病毒载体将葡萄糖脑苷酶(GC)基因转移到正常小鼠和人类造血干细胞，或者在腺相关病毒载体转移到肌肉细胞，并成功地将转导的小鼠细胞移植到小鼠体内，但迄今为止进行的三项临床试验要么基因转移水平低，要么输注后植入失败，但只有一个例外。利用逆转录病毒、慢病毒和其他载体成功地将基因转移到神经元。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植入。我们假设，通过逆转录病毒和慢病毒载体转导的造血祖细胞或神经干细胞移植含有人葡萄糖脑苷酶的cDNAs。本研究将验证这样一种假设，即通过逆转录病毒和慢病毒载体转导的造血祖细胞或神经祖细胞移植到受体Gaucher L444P突变小鼠体内，并导致酶水平的提高。逆转录病毒和慢病毒载体都成功地证明了基因转移到神经元，并在受体动物的大脑中植入了转导的神经元和造血祖细胞。然而，尽管在动物模型中取得了这种成功，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。为了实现这个目标，我们将：1)更完整地描述新建立的Gaucher L444P突变小鼠的特性；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提高程度；3)鞘内注射体外转导的造血祖细胞或神经干细胞，以确定这些细胞是否会在脑内重新繁殖并表达葡萄糖脑苷酶活性。在这项研究完成时，我们将确定慢病毒介导的人GC基因转移是否会导致受体Gaucher突变小鼠组织中的酶水平持续和改善，特别是大脑中的酶水平。

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.