ANIMAL MODEL OF GLOBOID CELL LEUKODYSTROPHY IN RHESUS: KRABBES DISEASE

恒河猴球状细胞脑白质营养不良的动物模型：克拉布斯病

• 批准号: 6277433
• 负责人: GARY B BASKIN
• 金额: $ 10.02万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 1999-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6277433
• 关键词: Mammalia; Primates; congenital disorders; gene therapy; genetics; model design /development; nervous system; orphan disease /drug; technology /technique

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

球状细胞白质营养不良是一种罕见的常染色体隐性遗传 由溶酶体B-半乳糖苷酶活性低水平引起的疾病 髓鞘代谢中重要的酶。一只幼年恒河猴 这种疾病是1989年在TRPRC被诊断出来的。在其间的九个月里 多年来，我们故意对这群动物进行近亲繁殖，并在1996年 观察了另外两名受影响的婴儿。这两个纯合子 受感染的动物使我们在大卫·温格博士的合作下， 费城杰斐逊医学院，以确定 恒河猴GALC基因的致病突变和明确的 用聚合酶链式反应鉴定22只携带动物。恒河猴GALC的序列 Cdna与人类的同源性为98%，推导出的氨基酸 序列与人类的序列有97%的同源性。对此负责的突变 对于这些猴子中的GLD，包括AC的缺失 对应于cDNA387和388位的二核苷酸。这 导致帧移位，导致过早终止密码子。GALC 对2名纯合子患者，21名正常和20名受试者进行了活性测定。 携带者猴子。两名受影响的婴儿GALC活性低于 正常的2%。21只正常猴子的平均活动量为0.94 Nmol/hr/mg蛋白质，而20个携带者的平均含量为0.53。 受影响的婴儿大脑中的精神激素水平非常高。 这些结果已经发表(Luzi P，Rafi MA，Victoria T，Baskin 英国队，温格地区检察官。恒河猴的特征 半乳糖脑苷酶(GALC)基因的克隆及鉴定 引起球状细胞白质营养不良(Krabbe病)的突变 灵长类。基因组学42：319-324,1997；Baskin GB，Ratterree M，Davison BB，Falkenstein KP，Clarke MR，英格兰JD，Vanier MT，Luzi P，Rafi MA， 温格检察官。遗传性半乳脑苷酶缺乏症(球状细胞 脑白质营养不良、Krabbe病)。(已提交)。我们 建立成纤维细胞和EBV转化的淋巴细胞培养体系 这些纯合子婴儿，并已开始体外治疗 对含有人GALC基因的逆转录病毒载体的研究。这 非人灵长类动物模型将在基因治疗的研究中具有价值 这种疾病和类似的疾病。支持这一点的赠款申请