ANIMAL MODEL OF GLOBOID CELL LEUKODYSTROPHY IN RHESUS MONKEYS: LYSOSOMAL STORAGE

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。 此后我们 故意近亲繁殖这组动物，并在1996年观察到两个 更多受影响的婴儿 这两只纯合子受影响的动物 让我们与杰斐逊医疗公司的大卫温格博士合作， 费城大学，以确定致病突变，在 恒河猴GALC基因，并明确确定22个载体动物， PCR法 恒河猴GALC cDNA序列与野生型GALC cDNA序列有98%的同源性。 推导的氨基酸序列与人的同源性为97%， 人类 在这些猴子中导致GLD的突变包括 对应于cDNA位置的AC二核苷酸的缺失 387和388。 这会导致帧移位，导致过早的 终止密码子 在2个受影响的纯合子中测量GALC活性， 21只正常猴和20只携带者猴。 2名受影响的婴儿患有GALC 低于正常值的2%。 21个正常人的平均活动 猴子为0.94 nmol/hr/mg蛋白质，而20 携带者为0.53。 受影响的大脑中的精神病水平 婴儿非常高。 我们建立了成纤维细胞和EB病毒转化的 从这些纯合子婴儿的淋巴细胞培养， 使用含有以下的逆转录病毒载体的体外治疗研究： 人GALC基因。 我们进行了绒毛取样， 出生季节，并准确地诊断了遗传状态， 胎儿出生前每年。 在一个相关的子项目中， 开始初步研究，以确定最适当的方式， 将治疗基因传递给受影响的猴子 这种非人灵长类动物 这一模型对研究此类肿瘤的基因治疗及类似疾病的基因治疗具有重要意义 紊乱 基金基础资助，风险研究出版物Luzi P， Rafi MA，维多利亚T，巴斯金GB，温格DA。 恒河猴半乳糖苷酶（GALC）cDNA和基因，和 引起球状细胞脑白质营养不良的突变的鉴定 （Krabbe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.遗传半乳糖苷酶 恒河猴缺乏症（球样细胞脑白质营养不良，克拉伯病） 猴子 第48章：你是谁？

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. Since then, 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. 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. We have performed chorionic villus sampling for 3 birth seasons and have accurately diagnosed the genetic status of the fetuses each year prior to birth. In a related subproject, we have begun preliminary studies to determine the most appropriate way to deliver therapeutic genes to affected monkeys. This nonhuman primate model will be valuable for studies of gene therapy of this and similar disorders. FUNDING Base Grant, Venture Research PUBLICATIONS 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. Lab Anim Sci 48:476-