Entrapment of retroviral vector producer cells in three-dimensional alginate scaffolds for potential use in cancer gene therapy.

Abstract

We explored the possibility of entrapping retroviral vector producing cells (VPC) within porous 3D matrix to induce a local and sustained release of viral particles to the malignant milieu. PA317/STK, which constantly shed retroviral vectors, was used to transduce cancer cells with the herpes simplex virus thymidine kinase (HSV-tk) gene. Once HSV-tk is expressed, it preferentially phosphorylates nucleoside analog prodrugs, such as ganciclovir (GCV) and N-methanocarbathymidine (N-MCT), to their active triphosphate metabolites, which when incorporated into cellular DNA cause cell death. PA317/STK cells were seeded within 3D alginate scaffold at two different cell densities via static seeding procedure. In vitro assays determined that PA317/STK seeded at high-cell density in scaffolds maintained constant cell number, low cell leakage, and spheroid morphology with viral vector transfection activity. Postcell-seeding viral vector activity was confirmed by transfection of murine colon cancer cells (MC38) with conditioned media originated from VPC-containing scaffolds and the subsequent ability to generate N-MCT triphosphate. Preliminary in vivo transplantation of VPC-containing scaffolds into the peritoneal cavity of mice bearing intraperitoneal MC38 tumors with 2 weeks subsequent GCV administration resulted in a significantly higher survival rate relative to control groups. Our results demonstrate the feasibility of employing alginate scaffolds to efficiently entrap and support PA317/STK cells for cancer gene therapy.

• Authors: Mona Dvir-Ginzberg, Alexander Konson, Smadar Cohen, Riad Agbaria
• Journal: Journal of biomedical materials research. Part B, Applied biomaterials (J Biomed Mater Res B Appl Biomater) Vol. 80 Issue 1 Pg. 59-66 (Jan 2007) ISSN: 1552-4973 [Print] United States
• PMID: 16680730 (Publication Type: Journal Article)
• Copyright: 2006 Wiley Periodicals, Inc.

Chemical References

• Alginates
• Antiviral Agents
• Hexuronic Acids
• Glucuronic Acid
• Thymidine Kinase
• (north)-methanocarbathymidine
• Ganciclovir
• Thymidine

Topics

• Alginates (chemistry)
• Animals
• Antiviral Agents (pharmacology)
• Cell Culture Techniques
• Cells, Immobilized (enzymology)
• Ganciclovir (pharmacology)
• Genetic Therapy
• Genetic Vectors
• Glucuronic Acid (chemistry)
• Herpes Simplex (enzymology, genetics)
• Hexuronic Acids (chemistry)
• Mice
• Neoplasms, Experimental (enzymology, genetics, therapy)
• Retroviridae
• Thymidine (analogs & derivatives, pharmacology)
• Thymidine Kinase (biosynthesis, genetics)
• Tumor Cells, Cultured