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。从那时起，我们 故意对这群动物进行近亲繁殖，并在1996年观察到两只 其他受影响的婴儿。这两只纯合子感染的动物 允许我们与大卫·温格博士合作，杰斐逊医疗 费城大学，以确定致病突变在 恒河猴GALC基因及明确鉴定22只携带该基因的动物 聚合酶链式反应。恒河猴GALC基因的序列与恒河猴的 推导的氨基酸序列与人的氨基酸序列同源性为97%。 人类。导致这些猴子GLD的突变包括 与cDNA位对应的AC二核苷酸的缺失 387和388。这会导致帧移动，从而导致过早 停止密码子。检测2例受累纯合子的GALC活性， 正常猴21只，携带猴20只。两名受影响的婴儿患有GALC 活动量低于正常的2%。21名正常人的平均活动量 猴子的蛋白质含量为0.94nmol/小时/毫克，而平均为20 携带者为0.53。精神分裂症患者脑组织中的精神药物水平 婴儿都长得很高。我们建立了成纤维细胞和EBV转化 来自这些纯合子婴儿的淋巴细胞培养，并已开始 重组逆转录病毒载体的体外治疗研究 人类GALC基因。我们已经做了3次绒毛取样。 出生季节和准确诊断的遗传状态 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，以确定最合适的方式 将治疗性基因传递给受影响的猴子。这种非人类的灵长类动物 该模型对这类疾病的基因治疗研究具有一定的参考价值 精神错乱。基金基础补助金，创业研究出版物鲁兹·P， 拉菲·马，维多利亚·T，巴斯金·英格，温格·达尔。刻画人物形象 猕猴半乳脑苷酶(GALC)基因和 球状细胞白质营养不良突变的鉴定 (Krabbe病)在这种灵长类动物身上。基因组学42：319-324,1997；Baskin GB，Ratterree M，Davison BB，Falkenstein KP，Clarke MR，英格兰JD， 瓦尼尔·MT，卢齐·P，拉菲·马，温格·达尔。遗传性半乳脑苷酶 恒河猴的缺乏症(球状细胞白质营养不良、Krabbe病) 猴子。实验室动画科学48：476-