ANIMAL MODEL OF GLOBOID CELL LEUKODYSTROPHY IN RHESUS: KRABBES DISEASE

球样细胞脑白质营养不良是一种罕见的常染色体隐性遗传病 由低水平的B-半乳糖苷酶活性引起的疾病， 髓鞘代谢中重要的酶。 一只幼年恒河猴 该疾病于1989年在TRPRC被诊断出。 在中间的九个 1996年，我们有意让这群动物近亲繁殖， 观察了另外两名受影响的婴儿。 这两个纯合子 受影响的动物让我们与大卫温格博士合作， 杰斐逊医学院，费城，以确定 恒河猴GALC基因的致病突变， 用PCR方法对22只带菌者进行了鉴定。 恒河猴GALC的序列 cDNA序列与人的同源性为98%，推导的氨基酸序列与人的同源性为98%。 基因序列与人类有97%的相似性 突变导致了 对于这些猴子中的GLD， 对应于cDNA位置387和388的二核苷酸。 这 导致移码，导致提前终止密码子。 GALC 在2例受影响的纯合子、21例正常人和20例 携带病毒的猴子 2名受影响的婴儿的GALC活性低于 正常的2%。 21只正常猴的平均活性为0.94 nmol/hr/mg蛋白质，而20名携带者的平均值为0.53。 受影响的婴儿大脑中的Psychosine水平非常高。 这些结果已经发表（Luzi P，Rafi MA，维多利亚T，巴斯金 GB，Wenger DA.恒河猴的特征 半乳糖苷酶（GALC）cDNA和基因，以及鉴定 突变导致球样细胞脑白质营养不良（克拉伯病），在这个 灵长类动物 Genomics 42：319-324，1997;巴斯金GB，Ratterree M，Davison BB，Falkenstein KP，Clarke MR，England JD，Vanier MT，Luzi P，Rafi MA， Wenger DA.遗传性半乳糖苷酶缺乏症（球状细胞 脑白质营养不良、克拉布病）。 （已提交）。 我们 建立了成纤维细胞和EBV转化的淋巴细胞培养物， 这些纯合子婴儿，并已开始在体外治疗 使用含有人GALC基因的逆转录病毒载体进行的研究。 这 非人灵长类动物模型将对研究基因治疗有重要价值。 以及类似的疾病。 一份资助申请

Globoid cell leukodystrophy is a rare autosomal recessive genetic disease caused by low levels of B-galactosidase activity, a lysosomal enzyme important in myelin metabolism. An infant rhesus monkey with this disease was diagnosed at TRPRC in 1989. In the intervening nine years, we intentionally inbred this group of animals and in 1996 observed two additional affected infants. These two homozygous affected animals allowed us, in collaboration with Dr. David Wenger, Jefferson Medical College, Philadelphia, to identify the disease-causing mutation in the rhesus GALC gene and to unequivocally identify 22 carrier animals by PCR. The sequence of the rhesus GALC cDNA is 98% identical to the human, and the deduced amino acid sequence is 97% identical to that of humans. The mutation responsible for GLD in these monkeys consists of the deletion of the AC dinucleotide corresponding to cDNA positions 387 and 388. This results in a frame shift, leading to a premature stop codon. GALC activity was measured in the 2 homozygous affected, 21 normal and 20 carrier monkeys. The 2 affected infants had a GALC activity less than 2% of normal. The average activity for 21 normal monkeys was 0.94 nmol/hr/mg of protein, while the average for 20 carriers was 0.53. Psychosine levels in the brains of affected infants were very high. These results have been published (Luzi P, Rafi MA, Victoria T, Baskin GB, Wenger DA. Characterization of the rhesus monkey galactocerebrosidase (GALC) cDNA and gene, and identification of the mutation causing globoid cell leukodystrophy (Krabbe disease) in this primate. Genomics 42:319-324, 1997; Baskin GB, Ratterree M, Davison BB, Falkenstein KP, Clarke MR, England JD, Vanier MT, Luzi P, Rafi MA, Wenger DA. Genetic Galactocerebrosidase deficiency (globoid cell leukodystrophy, Krabbe disease) in rhesus monkeys. (SUBMITTED). We established fibroblast and EBV-transformed lymphocyte cultures from these homozygous infants, and have initiated in vitro therapeutic studies with retroviral vectors containing the human GALC gene. This nonhuman primate model will be valuable for studies of gene therapy of this and similar disorders. A grant application to support this