Studies for Gene Therapy of Sphingolipidosis

Gene analysis of Japanese lipidodes patients ：We analyzed gene from various Japanese patients with lipidoses and found that the distribution of gene mutations was quite different from that from other ethnic groups.Gene therapy ：The deficiency of human？glucuronidase（HBG） results in MPS type VII（Sly syndrome）。在这一研究中，我们测试了CD 18/CD 11b阳性细胞用于murine MPS VII的基因治疗的能力。我们从合成正常老鼠中收获了骨头marrow细胞，并在2周内含有CSF-1的介质中被培养。超过95%的细胞对CD 18/CD 11b的双重阳性。We gave 2×10-D16-D1 cells to the non-myeloablated Sly mouse。一周后移植，捐赠者细胞流行的肝脏和繁殖。HBG活动从0.9±0.7至28.4± 12.5 u/mg和0.7±0.4至29.7± 23.1 u/mg分别在利害相关地增加。哇，病理学已经改变了。HBG在Liver和spleen decreased B5周至3.7±1.5和2.3±0.5的反应 ... More 是的，但病理学改进是有意义的，而且糖胺聚糖的储存是相当清楚的。下一步，CD 18/CD 11b细胞是由上述方法收集的，由HBG表达逆转录病毒（MFG-HBG）转换的，并给予非骨髓分裂的斯利鼠标。在移植后5周内， HBG活动仅在控制水平上边缘。However，许多HBG积极的细胞被观察到，病理学的改善是显著的。这些数据推荐CD 18/CD 11b细胞移植是为了治疗murine MPS VII而没有myeloablation，并且CD 18/CD 11b细胞可能是用于基因治疗的良好目标。基因转移到神经元细胞：Because significant prenatal pathology occurs in genetic diseases， postnatal therapy is often not sufficient to correct them， especially if dame has occurred in the central nervous system。在这种疾病中，需要进行全面治疗。Adenovirus介导的基因转移可能是有用的，因为这种目的是由于它的广泛宿主范围和有效的基因转移到不同的器官。However，大多数基因转移研究都被带出了一个相对较晚的胚胎发育阶段。在这项研究中，一种重新组合的腺病毒表达了E. coli lacZ gene （AxCALacZ） was administered into the rat embryos at the 8 th to 12 th day of gestation。这项研究的结果揭示了LacZ基因被描述为许多不同的器官，包括肝脏、心脏、皮肤和大脑。这些结果建议，即腺病毒载体可能对许多遗传疾病的原始基因疗法是有用的。Less（低）

Gene analysis of Japanese lipidodes patients :We analyzed gene from various Japanese patients with lipidoses and found that the distribution of gene mutations was quite different from that from other ethnic groups.Gene therapy :The deficiency of human β-glucuronidase(HBG) results in MPS type VII(Sly syndrome). In this study, we tested the ability to target CD18/CD11b positive cells with gene therapy for murine MPS VII. We harvested bone marrow cells from syngeneic normal mice and cultivated in CSF-1 containing medium for 2 weeks. More than 95% of the cells were double positive for CD18/CD11b by flowcytometry. We gave 2×10ィイD16ィエD1 cells to the non-myeloablated Sly mouse. One week post-transplantation, donor cells populated liver and spleen. The HBG activity increased from 0.9±0.7 to 28.4±12.5u/mg and 0.7±0.4 to 29.7±23.1u/mg in liver and spleen respectively. However, the pathology was unchanged. The HBG activity in liver and spleen decreased b 5 weeks to 3.7±1.5 and 2.3±0.5 respectivel … More y, but the pathological improvements were significant and glycosaminoglycan storage was largely cleared. Next, the CD18/CD11b cells were collected from Sly mouse by the method above, transduced by HBG expressing retrovirus(MFG-HBG), and given to non-myeloablated Sly mouse. By 5 weeks post-transplantation, HBG activity was only marginally above the control level. However, many HBG positive cells were observed and pathological improvement was significant. These data suggest that CD18/CD11b cells transplantation is promising for treatment of murine MPS VII without myeloablation, and CD18/CD11b cells may be a good targets for gene therapy for Sly syndrome.Gene transfer to neuronal cells :Because significant prenatal pathology occurs in genetic diseases, postnatal therapy is often not sufficient to correct them, especially if damage has occurred in the central nervous system. In such diseases, prenatal treatment will be required. Adenovirus mediated gene transfer may be useful for this purpose because of its wide host range and efficient gene transfer to various organs. However, most of the gene transfer studies have been carried out at a relatively late stage of embryogenesis. In this study, a recombinant adenovirus expressing the E. coli lacZ gene (AxCALacZ) was administered into the rat embryos at the 8th to 12th day of gestation. The results of the study revealed that the lacZ gene was expressed in many different organs including liver, heart, skin and brain. These results suggest that the adenovirus vector may be useful for the prenatal gene therapy of many genetic diseases. Less