Selection of genetically modified hematopoietic cells in vitro and in vivo using alkylating agent lysomustine
| Autor: | Galina L. Levit, Tatyana Grinenko, F. N. Rozov, Victor P. Krasnov, A.V. Belyavsky |
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| Rok vydání: | 2010 |
| Předmět: |
Alkylating Agents
Genetic enhancement Genetic Vectors Biophysics Bone Marrow Cells Biology Transfection Biochemistry CXCR4 Nitrosourea Compounds Viral vector Mice O(6)-Methylguanine-DNA Methyltransferase Immune system Cell Line Tumor medicine Animals Humans Cytotoxic T cell Molecular Biology Bone Marrow Transplantation Lysine Lentivirus Cell Biology Carmustine Molecular biology Mice Inbred C57BL medicine.anatomical_structure Cancer research Female Bone marrow Stem cell |
| Zdroj: | Analytical Biochemistry. 404:149-154 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/j.ab.2010.04.037 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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