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Targeted gene transfer into head and neck squamous cell carcinoma by nanosecond pulsed laser-induced stress waves
Koji Araki1, Daisuke Mizokami, Nobuaki Tanaka
1Department of Otolaryngology-Head and Neck Surgery, National Defense Medical College, 3-2 Namiki, Tokorozawa, Saitama, 359-8513, Japan, kojaraki@ndmc.ac.jp.
Abstract:
The search for new strategies to improve patient outcome and organ preservation is of great clinical interest in head and neck squamous cell carcinoma (HNSCC) treatment, and gene therapy is expected to play a promising role. In this study, we demonstrated the value of laser-induced stress waves (LISWs) as a novel method for nonviral gene transfer for gene therapy in HNSCC. The in vitro and in vivo transfection efficiency as well as in vitro cytotoxicity for HNSCC was investigated. Green fluorescent protein (GFP) expression and cell viability were analyzed in vitro after administration of a GFP-expressing plasmid or cationically modified GFP-expressing plasmid with LISW application. Luciferase expression in xenograft tumors was also quantitatively analyzed in vivo. The GFP gene was successfully transfected into HNSCC cells in vitro by LISW application. The cationically modified plasmid demonstrated enhanced transfection efficiency. LISWs are not associated with adverse effects after application to cells in vitro. The reporter genes were also successfully transfected into HNSCC tumors in vivo by LISW application. This technique is site specific, safe, and easily applicable for practical purposes. LISW gene therapy with therapeutic factors that inhibit tumor growth therefore has the potential as a future treatment for HNSCC.
Insights
Laser-induced stress waves (LISWs) offer a novel, safe method for nonviral gene transfer in head and neck squamous cell carcinoma (HNSCC) gene therapy. This technique successfully transfected genes both in vitro and in vivo, showing potential for future HNSCC treatments.
Area of Science:
- Oncology
- Gene Therapy
- Biomedical Engineering
Background:
- Improving patient outcomes and organ preservation in head and neck squamous cell carcinoma (HNSCC) treatment is a clinical priority.
- Gene therapy presents a promising avenue for novel HNSCC treatment strategies.
- Nonviral gene transfer methods are crucial for safe and effective gene delivery.
Purpose of the Study:
- To evaluate laser-induced stress waves (LISWs) as a novel nonviral gene transfer method for HNSCC.
- To assess the in vitro and in vivo transfection efficiency of LISWs in HNSCC.
- To investigate the in vitro cytotoxicity of LISWs in HNSCC.
Main Methods:
- In vitro and in vivo transfection of HNSCC cells and xenograft tumors using Green Fluorescent Protein (GFP) and luciferase reporter genes.
- Application of LISWs for gene delivery with standard and cationically modified plasmids.
- Analysis of GFP expression and cell viability in vitro, and luciferase expression in vivo.
Main Results:
- Successful in vitro transfection of HNSCC cells with GFP gene using LISWs.
- Cationically modified plasmids demonstrated enhanced transfection efficiency.
- LISWs showed no adverse effects on cell viability in vitro.
- Successful in vivo transfection of reporter genes into HNSCC xenograft tumors.
Conclusions:
- LISW application is a safe, site-specific, and practical method for nonviral gene transfer in HNSCC.
- LISW-mediated gene therapy holds potential for future HNSCC treatment, particularly with therapeutic factors inhibiting tumor growth.
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