Hubel A, Stroncek D, Pan D, Whitley C B, McCullough J
Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis 55455, USA.
J Hematother. 1998 Dec;7(6):505-14. doi: 10.1089/scd.1.1998.7.505.
Mucopolysaccharidosis type I (MPS I) results from a deficiency of alpha-L-iduronidase enzyme (IDUA), an enzyme responsible for the catabolism of glycosaminoglycans. Genetically modified progenitor cells may permit a therapeutic effect similar to that obtained from allogeneic BMT without the associated risks. To that end, CD34+ peripheral blood hematopoietic progenitor cells from patients with MPS I were mobilized using G-CSF, collected by apheresis, and enriched using avidin-biotin separation techniques. These cells were cultured in a hollow fiber bioreactor and transduced with a retroviral vector (LP1CD) containing the cDNA for human IDUA and a murine dihydrofolate reductase (DHFR) enzyme. Approximately 4%-16% of the colonies expressed methotrexate drug resistance. Expression of the IDUA enzyme in the progenitor cells was initially high and declined after approximately 10 days of culture. These results indicate that PBPC from patients with MPS I can be mobilized, isolated, enriched, and transduced with a therapeutic gene.
I型黏多糖贮积症（MPS I）是由于α-L-艾杜糖醛酸酶（IDUA）缺乏所致，该酶负责糖胺聚糖的分解代谢。基因改造的祖细胞可能产生与异基因骨髓移植相似的治疗效果，且无相关风险。为此，使用粒细胞集落刺激因子（G-CSF）动员I型黏多糖贮积症患者的CD34+外周血造血祖细胞，通过单采术收集，并采用抗生物素蛋白-生物素分离技术进行富集。将这些细胞在中空纤维生物反应器中培养，并用含有人类IDUA cDNA和小鼠二氢叶酸还原酶（DHFR）的逆转录病毒载体（LP1CD）进行转导。约4%-16%的集落表达甲氨蝶呤耐药性。祖细胞中IDUA酶的表达最初较高，培养约10天后下降。这些结果表明，I型黏多糖贮积症患者的外周血祖细胞可以被动员、分离、富集并用治疗性基因进行转导。
