S 45 Selection of genetically modified hematopoietic cells in vitro and in vivo using alkylating agent lysomustine [Text] / F. N. Rozov, T. S. Grinenko, G. L. Levit, V. P. Krasnov, A. V. Belyavskii // Analytical Biochemistry . - 2010. - Vol. 404, № 2. - P149-154 Рубрики: ХИМИЧЕСКИЕ НАУКИ Аннотация: 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 O6-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 |