Bosch A, Perret E, Desmaris N, Trono D, Heard J M
Unité de Rétrovirus et Transfert Génétique, CNRS URA 1930, Institut Pasteur, Paris, France.
Hum Gene Ther. 2000 May 20;11(8):1139-50. doi: 10.1089/10430340050015194.
Gene transfer vectors derived from human immunodeficiency virus (HIV-1) efficiently transduce nondividing cells and remain stably integrated in their genome. Long-term expression of reporter genes has been documented after intracerebral injection of these vectors. Using a HIV-based vector, we looked for a reversal of brain damage in the beta-glucuronidase-deficient mucopolysaccharidosis type VII mouse, an animal model of human lysosomal storage diseases. The vector suspension was injected stereotactically in the brain of 10-week-old animals, an age at which storage lesions are patent in glia, perivascular cells, and neurons. Either a single intrastriatal injection or multiple injections in both cerebral hemispheres and in the cerebellum were performed. Local tolerance, enzyme delivery, and correction of storage lesions were investigated by comprehensive analysis of serial sections of the entire brain of mice killed 6 or 16 weeks postinjection. Histochemical staining detected enzyme activity in widely distributed areas, the size of which increased with time. Clearance of lysosomal storage extended far beyond enzyme-positive areas. In mice receiving multiple injections of the vector, complete correction or significant reduction of the pathology was observed in every section, suggesting disease regression in the entire brain. These results may have implications for the treatment of neurological symptoms in lysosomal storage diseases.
源自人类免疫缺陷病毒(HIV-1)的基因转移载体能够有效地转导非分裂细胞,并稳定地整合到其基因组中。脑内注射这些载体后,已证明报告基因可长期表达。我们使用基于HIV的载体,在β-葡萄糖醛酸酶缺陷型VII型粘多糖贮积症小鼠(一种人类溶酶体贮积病的动物模型)中寻找脑损伤的逆转方法。将载体悬浮液立体定向注射到10周龄动物的脑中,这个年龄段时,神经胶质细胞、血管周围细胞和神经元中的贮积病变已很明显。进行了单次纹状体内注射或在两个大脑半球和小脑中多次注射。通过对注射后6周或16周处死的小鼠全脑连续切片的综合分析,研究了局部耐受性、酶递送和贮积病变的纠正情况。组织化学染色在广泛分布的区域检测到酶活性,其范围随时间增加。溶酶体贮积物的清除远远超出了酶阳性区域。在接受多次载体注射的小鼠中,每个切片均观察到病理完全纠正或显著减轻,表明整个大脑中的疾病有所消退。这些结果可能对溶酶体贮积病的神经症状治疗具有启示意义。