A Hubel, D Stroncek, D Pan, C B Whitley, J McCullough
{"title":"粘多糖病患者外周血祖细胞的动员和转导1。","authors":"A Hubel, D Stroncek, D Pan, C B Whitley, J McCullough","doi":"10.1089/scd.1.1998.7.505","DOIUrl":null,"url":null,"abstract":"<p><p>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.</p>","PeriodicalId":77208,"journal":{"name":"Journal of hematotherapy","volume":"7 6","pages":"505-14"},"PeriodicalIF":0.0000,"publicationDate":"1998-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/scd.1.1998.7.505","citationCount":"2","resultStr":"{\"title\":\"Mobilization and transduction of peripheral blood progenitor cells in patients with mucopolysaccharidosis I.\",\"authors\":\"A Hubel, D Stroncek, D Pan, C B Whitley, J McCullough\",\"doi\":\"10.1089/scd.1.1998.7.505\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>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.</p>\",\"PeriodicalId\":77208,\"journal\":{\"name\":\"Journal of hematotherapy\",\"volume\":\"7 6\",\"pages\":\"505-14\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1998-12-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1089/scd.1.1998.7.505\",\"citationCount\":\"2\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of hematotherapy\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1089/scd.1.1998.7.505\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of hematotherapy","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1089/scd.1.1998.7.505","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 2
摘要
粘多糖病I型(MPS I)是由α - l -糖醛酸酶(IDUA)缺乏引起的,这种酶负责糖胺聚糖的分解代谢。基因修饰的祖细胞可能具有与同种异体骨髓移植相似的治疗效果,而没有相关的风险。为此,使用G-CSF动员来自MPS I患者的CD34+外周血造血祖细胞,通过分离收集,并使用亲和素-生物素分离技术进行富集。这些细胞在中空纤维生物反应器中培养,用含有人IDUA cDNA和鼠二氢叶酸还原酶(DHFR)酶的逆转录病毒载体(LP1CD)转导。约4%-16%的菌落表达甲氨蝶呤耐药。IDUA酶在祖细胞中的表达最初是高的,在培养大约10天后下降。这些结果表明,来自MPS I患者的PBPC可以被动员、分离、富集,并用治疗基因转导。
Mobilization and transduction of peripheral blood progenitor cells in patients with mucopolysaccharidosis I.
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.