Suicide gene transduction sensitizes murine embryonic and human mesenchymal stem cells to ablation on demand - a fail-safe protection against cellular misbehavior

• Published in: Gene therapy Vol. 9; no. 14; pp. 955 - 962
• Main Authors: FAREED, M. U, MOOLTEN, F. L
• Format: Journal Article
• Language: English
• Published: Basingstoke Nature Publishing Group 01.07.2002
• Subjects: Ablation; Aminopterin; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Animals; Apoptosis - drug effects; Apoptosis - genetics; Applied cell therapy and gene therapy; Biological and medical sciences; Biotechnology; Cell Line; Embryo cells; Fundamental and applied biological sciences. Psychology; Ganciclovir; Ganciclovir - therapeutic use; Gene therapy; Genes; Genetic Therapy; Genetic Vectors - administration & dosage; Health. Pharmaceutical industry; Hematopoietic Stem Cell Transplantation; Hematopoietic Stem Cells - cytology; Hemopoiesis; Humans; Hypoxanthine; Industrial applications and implications. Economical aspects; Kinases; Medical sciences; Mesenchymal stem cells; Mesoderm - cytology; Mice; Prodrugs; Prodrugs - therapeutic use; Simplexvirus - enzymology; Stem cell transplantation; Stem cells; Stem Cells - cytology; Suicide; Suicide genes; Thymidine; Thymidine kinase; Thymidine Kinase - genetics; Transduction, Genetic - methods; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; Human; Cell therapy; Stem cell; Rodentia; Retroviridae; Prodrug; Suicide gene; Virus; Vertebrata; Mammalia; Cell line; Mouse; Embryonic cell; Transduction; Gene therapy; Protection
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/sj.gt.3301771

Stem cells and their progeny constitute a potential resource for replacing damaged tissues or supplying missing functions, but also pose a threat of aberrant behavior, including neoplastic growth or immunopathology. Suicide genes introduced into these cells before transplantation might provide a means of addressing this threat by permitting the ablation of the cells if they subsequently misbehave. Retroviral transduction of the E. coli gpt and herpes thymidine kinase (HSVtk) suicide genes was used to determine the degree to which stem cells could be sensitized to the prodrugs 6-thioxanthine (6TX) and ganciclovir (GCV) respectively, and whether this sensitivity could persist over many cell generations. The ES-E14TG2a murine embryonic stem cell line was rendered sensitive to quantitative ablation at prodrug concentrations well tolerated by untransduced cells (50 microM 6TX, 1 microg/ml GCV). The HSVtk gene also conferred GCV sensitivity on human mesenchymal stem cells and hematopoietic precursors derived from the murine cells, although ablation was not complete. Because ES-E14TG2a cells are deficient in the cellular enzyme HPRT, they are sensitive to hypoxanthine/aminopterin/thymidine (HAT). This property enhanced the persistence of chemosensitivity in gpt-transduced cells by permitting cells that lost 6TX sensitivity to be ablated with HAT.