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Published on: August 12, 2014
Isolated limb perfusion for local gene delivery: efficient and targeted adenovirus-mediated gene transfer into soft
W K de Roos1, J H de Wilt, M E van Der Kaaden
1Department of Surgical Oncology, University Hospital Rotterdam/Daniel den Hoed Cancer Center, Rotterdam, and IntroGene B. V., Leiden, The Netherlands.
Objective:
To evaluate the potential of isolated limb perfusion (ILP) for efficient and tumor-specific adenovirus-mediated gene transfer in sarcoma-bearing rats.
Summary Background Data:
A major concern in adenovirus-mediated gene therapy in cancer is the transfer of genes to organs other than the tumor, especially organs with a rapid cell turnover. Adjustment of the vector delivery route might be an option creating tumor specificity in therapeutic gene expression.
Methods:
Rat hind limb sarcomas (5-10 mm) were transfected with recombinant adenoviruses. Intratumoral luciferase expression after ILP was compared with systemic administration, regional infusion, or intratumoral injection using a similar dose of adenoviruses carrying the luciferase marker gene. Localization studies using lacZ as a marker gene were performed to evaluate the intratumoral distribution of transfected cells after both ILP and intratumoral injection.
Results:
Intratumoral luciferase activity after ILP or intratumoral administration was significantly higher compared with regional infusion or systemic administration. After ILP, luciferase gene expression was minimal in extratumoral organs, whether outside or inside the isolated circuit. Localization studies demonstrated that transfection was confined to tumor cells lying along the needle track after intratumoral injection, whereas after ILP, lacZ expression was found in viable tumor cells and in the tumor-associated vasculature.
Conclusions:
Using ILP, efficient and tumor-specific gene transfection can be achieved. The ILP technique might be useful for the delivery of recombinant adenoviruses carrying therapeutic gene constructs to enhance tumor control.
Insights
Isolated limb perfusion (ILP) enables efficient and tumor-specific gene transfer using adenoviruses in sarcoma models. This method minimizes off-target gene expression in other organs, enhancing therapeutic potential.
Area of Science:
- Oncolytic virotherapy
- Gene therapy delivery systems
- Cancer research models
Background:
- Adenovirus-mediated gene therapy faces challenges with off-target gene transfer to healthy organs.
- Targeting organs with rapid cell turnover is a significant hurdle in cancer gene therapy.
- Optimizing vector delivery routes is crucial for achieving tumor-specific gene expression.
Purpose of the Study:
- To assess isolated limb perfusion (ILP) for effective and tumor-specific adenovirus gene transfer in rat sarcoma models.
- To compare ILP with other administration routes for gene delivery efficiency and specificity.
- To evaluate the distribution of gene transfection within tumors using ILP.
Main Methods:
- Rat hind limb sarcomas were transfected with recombinant adenoviruses.
- Intratumoral luciferase expression via ILP was compared to systemic, regional, or intratumoral delivery.
- Localization studies using lacZ marker gene assessed transfected cell distribution.
Main Results:
- ILP and intratumoral administration yielded significantly higher luciferase expression than regional or systemic routes.
- ILP resulted in minimal luciferase gene expression in extratumoral organs.
- ILP facilitated lacZ expression in tumor cells and associated vasculature, unlike needle-track confinement with intratumoral injection.
Conclusions:
- Isolated limb perfusion (ILP) achieves efficient and tumor-specific adenovirus-mediated gene transfection.
- The ILP technique shows promise for delivering therapeutic gene constructs to enhance tumor control.
- ILP offers a potential strategy to improve the safety and efficacy of adenovirus-based cancer gene therapy.
