Roman M, Axelrod J H, Dai Y, Naviaux R K, Friedmann T, Verma I M
Department of Pediatrics, University of California School of Medicine, San Diego, La Jolla 92093-0634.
Somat Cell Mol Genet. 1992 May;18(3):247-58. doi: 10.1007/BF01233861.
We have used retroviral vectors to introduce human or canine factor IX cDNAs into cultured primary murine and canine myoblasts and into the established murine myoblast cell line C2C12. In all cases, the stably infected cells produced biologically active factor IX in culture and secreted detectable amounts into the culture medium both before and after differentiation of the cells into myotubes. Myoblasts and differentiated myotubes are therefore capable of performing all the posttranslational modifications of the coagulation factor required for biological activity. We have grafted the genetically modified myoblasts into skeletal muscles of nude mice and have detected stable levels of circulating human factor IX for up to two months after grafting. We propose that grafting genetically modified primary myoblasts or established myoblast cell lines into skeletal muscle may represent a useful approach to gene therapy for a variety of genetic diseases, including intrinsic muscle disease and defects in circulating proteins as in the hemophilias.
我们已使用逆转录病毒载体将人或犬的凝血因子IX cDNA导入原代培养的小鼠和犬成肌细胞以及已建立的小鼠成肌细胞系C2C12中。在所有情况下，稳定感染的细胞在培养物中产生具有生物活性的凝血因子IX，并且在细胞分化为肌管之前和之后都向培养基中分泌可检测量的凝血因子IX。因此，成肌细胞和分化的肌管能够进行生物活性所需的凝血因子的所有翻译后修饰。我们已将基因修饰的成肌细胞移植到裸鼠的骨骼肌中，并在移植后长达两个月的时间内检测到循环人凝血因子IX的稳定水平。我们提出，将基因修饰的原代成肌细胞或已建立的成肌细胞系移植到骨骼肌中可能是治疗多种遗传疾病的有用方法，这些疾病包括内在性肌肉疾病以及如血友病中循环蛋白的缺陷。
