Selection of genetically modified hematopoietic cells in vitro and in vivo using alkylating agent lysomustine.
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.
Rozov
F N
FN
Engelhard Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, Moscow 119991, Russia.
Grinenko
T S
TS
Levit
G L
GL
Krasnov
V P
VP
Belyavsky
A V
AV
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
D000477:Alkylating Agents; D009607:Nitrosourea Compounds; C552857:lysomustine; D019853:O(6)-Methylguanine-DNA Methyltransferase; D008239:Lysine; D002330:Carmustine