Methods and compositions for enhancing viral and mRNA gene therapy with engineered RNA elements
Unmet Need
An outstanding challenge in creating effective gene therapies, particularly those based on adeno-associated virus (AAV) and mRNA delivery platforms, is overcoming sub-optimal transgene expression. Inefficiencies in translation and stability of therapeutic gene products limit therapeutic outcomes and necessitate high vector doses, increasing costs and safety risks. New strategies are required to enhance AAV transgene expression and stability to allow safer, more effective treatments at lower doses.
Technology
Duke inventors have developed a novel method to enhance both AAV and mRNA-based gene therapy by incorporating engineered flavivirus-derived RNA elements into recombinant constructs. This strategy leverages subgenomic flavivirus RNAs (sfRNAs), including compact dumbbell (DB) RNA structures, to improve transgene expression by stabilizing mRNA and promoting more efficient translation. In vitro studies demonstrated significantly higher transgene expression and prolonged mRNA half-life across multiple human cell lines (Huh7, HEK293, U87, RPE). In vivo studies in mice confirmed increased transgene expression in cardiac tissue without affecting AAV vector packaging efficiency or biodistribution. These findings provide strong preclinical evidence that this approach boosts therapeutic gene expression while preserving vector integrity, potentially enabling lower, safer dosing.
Other Applications
This technology could be applied to improve gene therapies targeting various tissues, as well as to enhance stability and expression of mRNA therapeutic platforms including vaccines and protein replacement therapy.
Advantages
- Increases transgene mRNA stability and translation efficiency in multiple cell types, indicating broadly conserved cellular mechanism.
- Demonstrated enhancement of AAV transgene expression in vivo in cardiac tissue.
- Applicable to both AAV and mRNA-based therapies, expanding potential therapeutic modalities.
- May enable lower therapeutic doses, reducing potential side effects and treatment costs.
- Compatible with existing AAV production systems, simplifying integration into current manufacturing pipelines.