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

恒河猴球状细胞脑白质营养不良的动物模型：溶酶体储存

• 批准号: 6116204
• 负责人: GARY B BASKIN
• 金额: $ 16.14万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6116204
• 关键词: Mammalia; congenital disorders; gene therapy; inborn metabolism disorder; 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. 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-

球形细胞白细胞营养不良是一种罕见的常染色体隐性遗传 由低水平的B-半乳糖苷酶活性（溶酶体）引起的疾病 酶在髓磷脂代谢中很重要。 一个婴儿的恒河猴与 这种疾病于1989年在TRPRC诊断出来。从那以后，我们 故意掩盖了这群动物，并在1996年观察到了两只 其他受影响的婴儿。 这两种纯合受影响的动物 允许我们与杰斐逊医疗博士合作 费城大学，以确定引起疾病的突变 恒河galc基因，并明确地识别22种载体动物 pcr。 恒河galc cDNA的序列与98％相同 人类，推导的氨基酸序列与97％相同 人类。 这些猴子中负责GLD的突变包括 与cDNA位置相对应的AC二核苷酸的缺失 387和388。这导致帧换档，导致过早 停止密码子。 GALC活性在受影响的2个纯合子中测量 21个正常和20个载体猴子。 受影响的婴儿有一个galc 活动少于正常的2％。 21正常的平均活动 猴子为0.94 nmol/hr/mg的蛋白质，而平均20 载体为0.53。 受影响的大脑的心理水平 婴儿很高。 我们建立了成纤维细胞和EBV转换 来自这些纯合婴儿的淋巴细胞培养物，并已开始 逆转录病毒载体的体外治疗研究 人galc基因。 我们已经进行了3 出生季节，并准确地诊断出 每年出生前的胎儿。 在一个相关的子项目中，我们有 开始初步研究，以确定最合适的方法 将治疗基因传递给受影响的猴子。 这个非人类的灵长类动物 模型对于研究基因治疗和类似的基因疗法很有价值 疾病。 资金基础赠款，风险研究出版物Luzi P， Rafi MA，Victoria T，Baskin GB，Wenger DA。 表征 恒河猴半乳脑脑化酶（Galc）cDNA和基因，以及 鉴定引起球形细胞白细胞营养不良的突变 （Krabbe病）在这种灵长类动物中。 基因组学42：319-324，1997;巴斯金 GB，Ratterree M，Davison BB，Falkenstein KP，Clarke MR，英格兰JD， Vanier MT，Luzi P，Rafi MA，Wenger DA。 基因半乳孢酶 恒河猴中的缺乏症（球形细胞白细胞营养不良，克拉布病） 猴子。 Lab Anim Sci 48：476-