Inhibition of miR-222 by Oncolytic Adenovirus-Encoded miRNA Sponges Promotes Viral Oncolysis and Elicits Antitumor Effects in Pancreatic Cancer Models
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Cell Lines
2.2. Single Guide RNA Design
2.3. Generation of CRISPR KO Cell Lines
2.4. miRNA Sponge Generation
2.5. Adenovirus Generation and Titration
2.6. Cytotoxicity Assay
2.7. Viral Release Assay
2.8. Intracellular Viral DNA Quantification
2.9. Gene Expression Analysis
2.10. miRNA Expression Analysis
2.11. eGFP Detection and Quantification
2.12. In Vivo Antitumor Study
2.13. In Vivo Toxicity Assay
2.14. Identification of Putative Targets of hsa-miR-222-3p
2.15. Statistical Analysis
3. Results
3.1. OncomiRs Downregulation Has a Strong Impact on Adenoviral Oncolysis
3.2. miR-222 Sponges Engineered in the Oncolytic Adenovirus AdNuPAR-E-miR222-S Reduce miR-222 Levels and Regulate Target Genes That Facilitate Viral Production
3.3. Inhibition of miR-222 by miR-222 Sponges Displayed Enhanced In Vitro Cytotoxicity and Improved Antitumoral Activity
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Cell Line | AdNuPAR-E-Scramble (IFU ± SEM) | AdNuPAR-E-miR222-S (IFU ± SEM) |
---|---|---|
PANC-1 | 0.255 ± 0.035 | 0.096 ± 0.0124 |
MIA PaCa-2 | 0.758 ± 0.099 | 0.385 ± 0.108 |
CP-15 Luc | 36.110 ± 2.039 | 15.132 ± 2.674 |
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Raimondi, G.; Gea-Sorlí, S.; Otero-Mateo, M.; Fillat, C. Inhibition of miR-222 by Oncolytic Adenovirus-Encoded miRNA Sponges Promotes Viral Oncolysis and Elicits Antitumor Effects in Pancreatic Cancer Models. Cancers 2021, 13, 3233. https://doi.org/10.3390/cancers13133233
Raimondi G, Gea-Sorlí S, Otero-Mateo M, Fillat C. Inhibition of miR-222 by Oncolytic Adenovirus-Encoded miRNA Sponges Promotes Viral Oncolysis and Elicits Antitumor Effects in Pancreatic Cancer Models. Cancers. 2021; 13(13):3233. https://doi.org/10.3390/cancers13133233
Chicago/Turabian StyleRaimondi, Giulia, Sabrina Gea-Sorlí, Marc Otero-Mateo, and Cristina Fillat. 2021. "Inhibition of miR-222 by Oncolytic Adenovirus-Encoded miRNA Sponges Promotes Viral Oncolysis and Elicits Antitumor Effects in Pancreatic Cancer Models" Cancers 13, no. 13: 3233. https://doi.org/10.3390/cancers13133233