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Published on: March 12, 2018
Efficient nonviral gene therapy with FasL and Del1 fragments in mice
Hisataka Kitano1, Atsushi Mamiya, Shinichiro Kokubun
1Division of Dental Surgery, Nihon University School of Medicine, Tokyo, Japan.
Background:
The expression of FasL in cancer cells is currently being explored as a potential cancer therapy. Because high levels of FasL are necessary for effective treatment, current methods typically rely on the use of highly efficient viral vectors. However, because viral vector-based gene therapy is associated with certain risks, the development of effective nonviral routes for gene delivery would be useful. The present study aimed to improve FasL gene therapy with a nonviral vector by taking advantage of the E3 and C1 domains of Del1 protein, which induces apoptosis and localizes to the extracellular matrix.
Methods:
Mouse explanted tumors derived from a human oral squamous cell carcinoma cell line, SCCKN, were treated with plasmids encoding FasL (pFasL), E3C1 (pE3C1), and a fusion of FasL and E3C1 (pFasL-E3C1). The plasmids were injected locally every 7 days along with a transfection reagent, Jet-PEI (PolyPlus-transfection, San Marcos, CA, USA).
Results:
All mice treated with a negative control plasmid or pFasL died within 49 days. By contrast, 83% of mice treated with pFasL-E3C1 survived longer than 49 days. Histochemical studies revealed that the fusion protein is localized to the stroma and induces apoptosis in stromal cells and adjacent parenchymal cells.
Conclusions:
The results obtained in the present study suggest that the protein deposition-based approach described, which makes use of the E3 and C1 domains of Del1, could comprise a novel method for cancer gene therapy with nonviral vectors.
Insights
This study introduces a novel nonviral gene therapy for cancer using a FasL-Del1 fusion protein. This approach enhances survival by inducing apoptosis in tumor stroma and surrounding cells, offering a safer alternative to viral vectors.
Area of Science:
- Oncology
- Gene Therapy
- Biotechnology
Background:
- FasL expression in cancer cells is a promising therapeutic strategy.
- Current viral vector methods for FasL gene delivery carry risks.
- Nonviral vectors are needed for safer and effective FasL gene therapy.
Purpose of the Study:
- To develop an improved nonviral FasL gene therapy using Del1 protein domains.
- To leverage Del1's E3 and C1 domains for enhanced apoptosis induction and extracellular matrix localization.
Main Methods:
- Mouse oral squamous cell carcinoma (SCCKN) tumors were treated with plasmids encoding FasL, E3C1, or a FasL-E3C1 fusion.
- Plasmids were administered via local injection with Jet-PEI transfection reagent every 7 days.
Main Results:
- Mice treated with control or FasL plasmids had a median survival of 49 days.
- 83% of mice treated with the FasL-E3C1 fusion protein survived longer than 49 days.
- Histochemical analysis confirmed fusion protein localization to the stroma, inducing apoptosis in stromal and parenchymal cells.
Conclusions:
- The Del1 E3/C1 domain-based protein deposition approach offers a novel nonviral vector strategy for cancer gene therapy.
- This method shows potential for improved therapeutic outcomes in cancer treatment.
- Further research into nonviral gene delivery systems is warranted for clinical applications.
