title: Selection of genetically modified hematopoietic cells in vitro and in vivo using alkylating agent lysomustine.
abstract: Efficient gene transfer into hematopoietic stem cells is vital for the success of gene therapy of hematopoietic and immune system disorders. An in vivo selection system based on a mutant form of the O(6)-methylguanine-DNA-methyltransferase gene (MGMTm) is considered one of the more promising strategies for expansion of hematopoietic cells transduced with viral vectors. Here we demonstrate that MGMTm-expressing cells can be efficiently selected using lysomustine, a nitrosourea derivative of lysine. K562 and murine bone marrow cells expressing MGMTm are protected from the cytotoxic action of lysomustine in vitro. We also show in a murine model that MGMTm-transduced hematopoietic cells can be expanded in vivo on transplantation into sublethally irradiated recipients followed by lysomustine treatment. These results indicate that lysomustine can be used as a potent novel chemoselection drug applicable for gene therapy of hematopoietic and immune system disorders.
authors.affiliation: Engelhard Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, Moscow 119991, Russia.
authors.forename: F N
authors.initials: FN
authors.lastname: Rozov
authors.forename: T S
authors.initials: TS
authors.lastname: Grinenko
authors.forename: G L
authors.initials: GL
authors.lastname: Levit
authors.forename: V P
authors.initials: VP
authors.lastname: Krasnov
authors.forename: A V
authors.initials: AV
authors.lastname: Belyavsky
mesh_terms: D000477:Alkylating Agents / Q000737:chemistry / Q000494:pharmacology*; D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism*; D016026:Bone Marrow Transplantation; D002330:Carmustine / Q000737:chemistry / Q000494:pharmacology; D045744:Cell Line, Tumor; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009607:Nitrosourea Compounds / Q000737:chemistry / Q000494:pharmacology*; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000235:genetics; D014162:Transfection
chemical_list: D000477:Alkylating Agents; D009607:Nitrosourea Compounds; C552857:lysomustine; D019853:O(6)-Methylguanine-DNA Methyltransferase; D008239:Lysine; D002330:Carmustine