Wilson J M, Chowdhury N R, Grossman M, Wajsman R, Epstein A, Mulligan R C, Chowdhury J R
Department of Internal Medicine, Howard Hughes Medical Institute, University of Michigan, Ann Arbor 48109.
Proc Natl Acad Sci U S A. 1990 Nov;87(21):8437-41. doi: 10.1073/pnas.87.21.8437.
Familial hypercholesterolemia is an inherited disease in humans that is associated with coronary artery disease and is caused by a deficiency of the receptor that mediates the internalization of low density lipoprotein (LDL). We have used an animal model for familial hypercholesterolemia, the Watanabe heritable hyperlipidemic (WHHL) rabbit, to design a therapeutic approach for this disease, which attempts to correct the hepatic defect in LDL receptor expression. Hepatocytes were harvested from WHHL rabbits, plated in primary cultures, and exposed to recombinant retroviruses capable of efficiently transferring a functional human LDL receptor gene. Genetically modified cells were harvested and infused into the portal vein of WHHL recipients, who were analyzed for metabolic consequences of human LDL receptor expression. Each animal exhibited a statistically significant decrease in total serum cholesterol 2-6 days after transplantation, with an eventual return to pretreatment levels. Proviral DNA sequences and virus-directed transcripts were detected in liver tissue 24 hr after transplantation. In situ hybridization demonstrated provirus expression in a small population of hepatocytes distributed in periportal sections of the liver. This study illustrates the potential of somatic gene therapy in ameliorating hyperlipidemia associated with familial hypercholesterolemia.
家族性高胆固醇血症是一种人类遗传性疾病，与冠状动脉疾病相关，由介导低密度脂蛋白（LDL）内化的受体缺陷引起。我们利用家族性高胆固醇血症的动物模型——渡边遗传性高脂血症（WHHL）兔，设计了一种针对该疾病的治疗方法，旨在纠正肝脏中LDL受体表达的缺陷。从WHHL兔中获取肝细胞，接种于原代培养物中，并使其接触能够有效转移功能性人类LDL受体基因的重组逆转录病毒。收集经过基因改造的细胞，并将其注入WHHL受体的门静脉，然后分析人类LDL受体表达的代谢后果。每只动物在移植后2 - 6天血清总胆固醇均出现统计学上的显著下降，最终又恢复到预处理水平。移植后24小时在肝脏组织中检测到前病毒DNA序列和病毒导向的转录本。原位杂交显示前病毒在分布于肝门周区域的一小部分肝细胞中表达。这项研究说明了体细胞基因治疗在改善与家族性高胆固醇血症相关的高脂血症方面的潜力。
