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Cell cycle independent infection and gene transfer by recombinant Sendai viruses
Sorin Armeanu1, Guy Ungerechts, Christian Bernloehr
1Internal Medicine I, University Clinic Tübingen, D-72076, Tübingen, Germany.
Abstract:
A common problem for viral vectors in the field of somatic gene therapy is the dependence of an efficient cellular transduction on the cell cycle phase of target cells. An optimized viral vector system should therefore transduce cells in different cell cycle phases equally to improve transduction efficiencies. Recent observations that recombinant Sendai viruses (SeV) can infect a broad range of different tissues suggested SeV to be a good candidate for future gene therapeutic strategies in which dividing and non-dividing cells have to be reached. However, detailed data on the influence of distinct cell cycle phases on the infection of SeV or related viruses are missing. We report that synchronization of NIH 3T3 cells as well as contact inhibition of human fibroblast cells did not exhibit any negative influence on SeV infection rates. Furthermore, different attractive target tissues like human umbilical cord derived cells or primary human hepatocytes can be reached by SeV efficiently. As an important information for further cell cycle studies of paramyxoviruses we discovered surprisingly that the DNA polymerase inhibitor aphidicolin (induces a G(1)/M arrest) functions as an inhibitor of SeV but not of an adenoviral expression vector. In conclusion, the results demonstrate SeV based vector particles to be an ideal tool to reach equally cells coexisting in different cell cycle phases.
Insights
Sendai viruses (SeV) efficiently infect cells regardless of their cell cycle phase, making them ideal for gene therapy. This viral vector system overcomes limitations of other vectors dependent on cell division for transduction.
Area of Science:
- Virology
- Gene Therapy
- Cell Biology
Background:
- Viral vector efficiency in somatic gene therapy often depends on the target cell's cycle phase.
- Optimized viral vectors should transduce cells in all cell cycle phases equally.
- Recombinant Sendai viruses (SeV) show potential for gene therapy due to broad tissue tropism.
Purpose of the Study:
- To investigate the influence of cell cycle phases on Sendai virus (SeV) infection efficiency.
- To evaluate SeV as a potential gene therapy vector for targeting cells in various cycle phases.
- To compare SeV's cell cycle independence with other viral vectors.
Main Methods:
- Synchronization of NIH 3T3 cells and contact inhibition of human fibroblasts were used to study cell cycle effects.
- SeV infection rates were assessed in synchronized and non-synchronized cells.
- Infection efficiency in target tissues (human umbilical cord cells, primary human hepatocytes) was evaluated.
- The effect of aphidicolin (a DNA polymerase inhibitor causing G1/M arrest) on SeV and adenoviral vector infection was tested.
Main Results:
- SeV infection rates were not negatively influenced by cell synchronization or contact inhibition.
- SeV efficiently infected human umbilical cord derived cells and primary human hepatocytes.
- Aphidicolin inhibited SeV infection but not adenoviral vector infection, suggesting a unique interaction with SeV replication.
Conclusions:
- SeV based vector particles demonstrate equal transduction efficiency across different cell cycle phases.
- SeV is a promising candidate for gene therapy applications requiring broad cell targeting.
- The differential effect of aphidicolin provides new insights into paramyxovirus cell cycle interactions.