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Updated: Jun 21, 2026
Expression of Transgenes in Native Bladder Urothelium Using Adenovirus-Mediated Transduction
Published on: October 6, 2022
Polymer-enhanced adenoviral transduction of CAR-negative bladder cancer cells
Laura M Kasman1, Sutapa Barua, Ping Lu
1Department of Microbiology, Medical University of South Carolina, Charleston, South Carolina, USA.
Abstract:
The application of adenoviral gene therapy for cancer is limited by immune clearance of the virus as well as poor transduction efficiency, since the protein used for viral entry (CAR) serves physiological functions in adhesion and is frequently decreased among cancer cells. Cationic polymers have been used to enhance adenoviral gene delivery, but novel polymers with low toxicity are needed to realize this approach. We recently identified polymers that were characterized by high transfection efficiency of plasmid DNA and a low toxicity profile. In this study we evaluated the novel cationic polymer EGDE-3,3' for its potential to increase adenoviral transduction of the CAR-negative bladder cancer cell line TCCSUP. The amount of adenovirus required to transduce 50-60% of the cells was reduced 100-fold when Ad.GFP was preincubated with the EGDE-3,3' polymer. Polyethyleneimine (pEI), a positively charged polymer currently used as a standard for enhancing adenoviral transduction, also increased infectivity, but transgene expression was consistently higher with EGDE-3,3'. In addition, EGDE-3,3'-supplemented transduction of an adenovirus expressing an apoptosis inducing transgene, Ad.GFP-TRAIL, significantly enhanced the amount of cell death. Thus, our results indicate that novel biocompatible polymers may be useful in improving the delivery of adenoviral gene therapy.
Insights
Novel cationic polymers enhance adenoviral gene therapy by improving viral delivery to cancer cells. EGDE-3,3
Area of Science:
- Biotechnology
- Molecular Biology
- Oncology
Background:
- Adenoviral gene therapy for cancer faces challenges including immune clearance and low transduction efficiency, particularly in CAR-negative cancer cells.
- Cationic polymers can enhance adenoviral gene delivery, but low-toxicity alternatives are needed.
- Previous research identified novel polymers with high plasmid DNA transfection efficiency and low toxicity.
Purpose of the Study:
- To evaluate the novel cationic polymer EGDE-3,3' for its ability to enhance adenoviral transduction in CAR-negative bladder cancer cells (TCCSUP).
Main Methods:
- Adenovirus expressing GFP (Ad.GFP) was preincubated with EGDE-3,3' polymer.
- Adenoviral transduction efficiency was assessed in TCCSUP cells.
- Comparison of EGDE-3,3' with polyethyleneimine (pEI) for enhancing adenoviral transduction.
- Evaluation of EGDE-3,3'-enhanced transduction of an adenovirus expressing a pro-apoptotic transgene (Ad.GFP-TRAIL).
Main Results:
- EGDE-3,3' reduced the required amount of adenovirus by 100-fold for 50-60% cell transduction.
- Transgene expression was consistently higher with EGDE-3,3' compared to pEI.
- EGDE-3,3'-supplemented transduction with Ad.GFP-TRAIL significantly increased cancer cell death.
Conclusions:
- Novel biocompatible polymers like EGDE-3,3' show significant potential for improving adenoviral gene delivery in cancer therapy.
- EGDE-3,3' offers a promising strategy to overcome limitations of adenoviral gene therapy, enhancing transduction efficiency and therapeutic outcomes.
