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Virus Delivery of CRISPR Guides to the Murine Prostate for Gene Alteration
Published on: April 27, 2018
Vector-based RNA interference against vascular endothelial growth factor-A significantly limits vascularization and
Francesca Wannenes1, Silvia Anna Ciafré, Francesco Niola
1Department of Experimental Medicine and Biochemical Sciences, University of Rome Tor Vergata, Via Montpellier 1, 00133 Rome, Italy.
Abstract:
RNA interference technology is emerging as a very potent tool to obtain a cellular knockdown of a desired gene. In this work we used vector-based RNA interference to inhibit vascular endothelial growth factor (VEGF) expression in prostate cancer in vitro and in vivo. We demonstrated that transduction with a plasmid carrying a small interfering RNA targeting all isoforms of VEGF, dramatically impairs the expression of this growth factor in the human prostate cancer cell line PC3. As a consequence, PC3 cells loose their ability to induce one of the fundamental steps of angiogenesis, namely the formation of a tube-like network in vitro. Most importantly, our "therapeutic" vector is able to impair tumor growth rate and vascularization in vivo. We show that a single injection of naked plasmid in developing neoplastic mass significantly decreases microvessel density in an androgen-refractory prostate xenograft and is able to sustain a long-term slowing down of tumor growth. In conclusion, our results confirm the basic role of VEGF in the angiogenic development of prostate carcinoma, and suggest that the use of our vector-based RNA interference approach to inhibit angiogenesis could be an effective tool in view of future gene therapy applications for prostate cancer.
Insights
Vector-based RNA interference effectively inhibits vascular endothelial growth factor (VEGF) in prostate cancer cells. This approach reduces tumor growth and vascularization, showing promise for gene therapy.
Area of Science:
- Oncology
- Molecular Biology
- Gene Therapy
Background:
- Vascular Endothelial Growth Factor (VEGF) is crucial for angiogenesis in various cancers, including prostate cancer.
- RNA interference (RNAi) offers a method to specifically inhibit gene expression.
Purpose of the Study:
- To investigate the efficacy of vector-based RNA interference in inhibiting VEGF expression in prostate cancer.
- To evaluate the impact of VEGF inhibition on angiogenesis and tumor growth in vitro and in vivo.
Main Methods:
- Utilized a plasmid carrying small interfering RNA (siRNA) targeting VEGF isoforms.
- Transduced human prostate cancer cell line PC3 with the vector for in vitro studies.
- Administered naked plasmid injection directly into androgen-refractory prostate xenografts for in vivo studies.
Main Results:
- Transduction with the VEGF-targeting siRNA plasmid significantly reduced VEGF expression in PC3 cells.
- Inhibition of VEGF impaired the in vitro formation of tube-like networks, a key step in angiogenesis.
- In vivo, a single plasmid injection decreased microvessel density and sustained long-term tumor growth inhibition in prostate xenografts.
Conclusions:
- VEGF plays a fundamental role in the angiogenic development of prostate carcinoma.
- Vector-based RNA interference targeting VEGF is a promising strategy for inhibiting angiogenesis.
- This approach holds potential for future gene therapy applications in prostate cancer treatment.
