Coexpression of p21WAF1/CIP1 in adenovirus vector transfected human primary hepatocytes prevents apoptosis resulting in improved transgene expression
| Autor: | Gerhard Wolff, Bernd Dörken, E Wehnes, Axel Schumacher, Peter Neuhaus, Andreas K. Nuessler, V. Ruppert, Leonid Karawajew |
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| Rok vydání: | 2003 |
| Předmět: |
Cyclin-Dependent Kinase Inhibitor p21
Transgene Genetic enhancement Genetic Vectors Gene Expression Apoptosis Biology medicine.disease_cause Adenoviridae Cell Line Viral vector Mice Cyclins Gene expression Genetics medicine Animals Humans Transgenes Molecular Biology Cell Cycle Genetic transfer Genetic Therapy Transfection Cell cycle Molecular biology Mice Mutant Strains Cell biology Hepatocytes Molecular Medicine |
| Zdroj: | Gene Therapy. 10:668-677 |
| ISSN: | 1476-5462 0969-7128 |
| DOI: | 10.1038/sj.gt.3301864 |
| Popis: | Replication-deficient adenovirus (Ad vector) is one of the most effective gene transfer systems. However, its employment in human gene therapy trials is hampered by Ad vector associated cytotoxicity and induction of apoptosis of the infected cells. Here, we identify one underlying mechanism as uncoupling of S phase and mitosis of the cell cycle leading to apoptosis and decline of transgene expression. Moreover, we demonstrate a strategy to avoid Ad vector associated cytotoxicity and induction of apoptosis in human primary hepatocytes by coinfection of Ad vector carrying the cDNA of choice and the cell cycle regulator p21(WAF1/CIP1) (p21). In addition, animal experiments were performed using Ad vector directed coexpression of p21 and human alpha 1-antitrypsin. As serum analysis of alpha 1-antitrypsin after Ad vector mediated gene transfer to the liver of mice revealed, this strategy resulted also in the improvement of transgene expression by two orders of magnitude. These data suggest that coexpression of p21 and Ad vector carrying a therapeutic gene may be a promising strategy to avoid cytotoxicity and induction of apoptosis leading to improved safety in human gene therapy. |
| Databáze: | OpenAIRE |
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