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Paramyxoviruses for Tumor-targeted Immunomodulation: Design and Evaluation Ex Vivo
Published on: January 7, 2019
RNAi-Mediated Knockdown of Cottontail Rabbit Papillomavirus Oncogenes Using Low-Toxicity Lipopolyplexes as a Paradigm
Uzma Ali1,2, Michael Bette3, Ghazala Ambreen1,2
1Department of Pharmaceutics and Biopharmaceutics, Philipps-Universität Marburg, 35037 Marburg, Germany.
Abstract:
The cottontail rabbit papillomavirus (CRPV)-associated VX2 carcinoma of the New Zealand White rabbit serves as a model system for human papillomavirus (HPV)-associated head and neck squamous cell carcinomas (HNSCCs). The aim of this study was to evaluate the tumor-inhibiting effect of RNAi-mediated knockdown of the CRPV oncogenes, E6 and E7, using siRNA-loaded lipopolyplexes (LPPs). VX2-carcinoma-derived cells were cultured for up to 150 passages. In addition, CRPV E6 and E7 oncogenes were transiently expressed in COS-7 cells. Efficiency and safety of LPPs were evaluated in both VX2 cells and the COS-7 cell line. Both of these in vitro CRPV systems were validated and characterized by fluorescence microscopy, Western blot, and RT-qPCR. Efficient knockdown of CRPV E6 and E7 was achieved in VX2 cells and COS-7 cells pretransfected with CRPV E6 and E7 expression vectors. Knockdown of CRPV oncogenes in VX2 cells resulted in reduced viability, migration, and proliferation and led to a G0/G1 block in the cell cycle. CRPV E6 and E7 siRNA-loaded LPPs could represent promising therapeutic agents serving as a paradigm for the treatment of papillomavirus-positive cancers and could be of value for the treatment of CRPV-associated diseases in the rabbit such as papillomas and cancers of the skin.
Insights
RNA interference targeting cottontail rabbit papillomavirus (CRPV) oncogenes E6 and E7 effectively inhibited VX2 carcinoma cell growth. This RNAi-mediated therapy shows promise for treating papillomavirus-associated cancers.
Area of Science:
- Oncology
- Virology
- Biotechnology
Background:
- The VX2 carcinoma in rabbits models human papillomavirus (HPV)-associated head and neck squamous cell carcinomas (HNSCCs).
- Cottontail rabbit papillomavirus (CRPV) oncogenes E6 and E7 drive tumor development.
Purpose of the Study:
- To assess the tumor-inhibiting potential of RNA interference (RNAi) targeting CRPV E6 and E7 oncogenes.
- To evaluate siRNA-loaded lipopolyplexes (LPPs) as a therapeutic delivery system.
Main Methods:
- CRPV-associated VX2 carcinoma cells and COS-7 cells expressing CRPV E6/E7 were used.
- Lipopolyplexes (LPPs) loaded with small interfering RNA (siRNA) targeting CRPV E6/E7 were developed.
- Knockdown efficiency and cellular effects were analyzed using fluorescence microscopy, Western blot, and RT-qPCR.
Main Results:
- Efficient knockdown of CRPV E6 and E7 oncogenes was achieved in both cell lines.
- CRPV oncogene knockdown in VX2 cells reduced cell viability, migration, and proliferation.
- A G0/G1 cell cycle arrest was observed following oncogene knockdown.
Conclusions:
- siRNA-loaded LPPs targeting CRPV E6 and E7 oncogenes demonstrate significant anti-tumor effects.
- This approach offers a potential therapeutic strategy for CRPV-associated diseases and papillomavirus-positive cancers.
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