Abstract
Background & Aims
The therapeutic potential of protein and RNA-based gene therapies is currently stalled by a critical delivery bottleneck, particularly within the cardiovascular space. While viral vectors (AAVs) and lipid nanoparticles (LNPs) remain the clinical standards, their utility is restricted by systemic immunogenicity, dose-limiting toxicities, and rigid packaging constraints.
In contrast, extracellular vesicles (EVs) are a promising next-generation drug delivery platform due to their innate targeting abilities, low immunogenicity, and full biocompatibility. To enable EVs for the delivery of genetic medicine, we have developed deVINCI, a modularly engineered extracellular vesicles for enhanced delivery of proteins and CRISPR-based therapeutics.
Methodology
The deVINCI platform utilized modular scaffolds fused to Cre recombinase or ABE8e base editors to enable efficient cargo loading and release. VSV-G pseudotyped EVs were produced in HEK293FT cells and characterized according to MISEV2023 guidelines (NTA, TEM, and Western blot for CD9/CD63/TSG101), with Gag-Pol VLPs as a benchmark. Functional delivery was quantified via flow cytometry in stoplight reporter cells, measuring RFP-to-GFP switching (Cre) or ABE8e-mediated GFP restoration. In vivo, AlexaFluor-labeled deVINCI-EVs were delivered via intramyocardial injection in Ai9 mice to evaluate biodistribution and functional recombination by a previously reported method (Ilahibaks et al., 2023).
Results
The deVINCI platform was optimized through iterative engineering of lipid anchors, linkers, and enzymatic cleavage sites to ensure efficient cargo loading and functional intracellular delivery. Benchmarked against Gag-Pol loaded virus like particels (VLPs), deVINCI EVs achieved 91.8% ± 2.6% efficiency in Cre-loxP cells. When loaded with ABE8e-RNPs, deVINCI-EVs maintained high on-target A>G base editing (92.6% ± 3.0%), significantly outperforming VLP-mediated delivery (81.9%) at the highest dose. VLPs showed a dose-dependent decline in performance, likely due to cytotoxicity. In vivo, the deVINCI EVs achieved heart-selective delivery with robust DNA recombination in cardiac tissue and minimal off-target activity in peripheral organs, as confirmed by immunofluorescent analysis of tdTomato expression.
Conclusion
By integrating a modular architecture that enables high-capacity cargo loading and heart-selective delivery, the deVINCI technology establishes a scalable 'EV 2.0′ modular delivery platform for precision medicine.
The therapeutic potential of protein and RNA-based gene therapies is currently stalled by a critical delivery bottleneck, particularly within the cardiovascular space. While viral vectors (AAVs) and lipid nanoparticles (LNPs) remain the clinical standards, their utility is restricted by systemic immunogenicity, dose-limiting toxicities, and rigid packaging constraints.
In contrast, extracellular vesicles (EVs) are a promising next-generation drug delivery platform due to their innate targeting abilities, low immunogenicity, and full biocompatibility. To enable EVs for the delivery of genetic medicine, we have developed deVINCI, a modularly engineered extracellular vesicles for enhanced delivery of proteins and CRISPR-based therapeutics.
Methodology
The deVINCI platform utilized modular scaffolds fused to Cre recombinase or ABE8e base editors to enable efficient cargo loading and release. VSV-G pseudotyped EVs were produced in HEK293FT cells and characterized according to MISEV2023 guidelines (NTA, TEM, and Western blot for CD9/CD63/TSG101), with Gag-Pol VLPs as a benchmark. Functional delivery was quantified via flow cytometry in stoplight reporter cells, measuring RFP-to-GFP switching (Cre) or ABE8e-mediated GFP restoration. In vivo, AlexaFluor-labeled deVINCI-EVs were delivered via intramyocardial injection in Ai9 mice to evaluate biodistribution and functional recombination by a previously reported method (Ilahibaks et al., 2023).
Results
The deVINCI platform was optimized through iterative engineering of lipid anchors, linkers, and enzymatic cleavage sites to ensure efficient cargo loading and functional intracellular delivery. Benchmarked against Gag-Pol loaded virus like particels (VLPs), deVINCI EVs achieved 91.8% ± 2.6% efficiency in Cre-loxP cells. When loaded with ABE8e-RNPs, deVINCI-EVs maintained high on-target A>G base editing (92.6% ± 3.0%), significantly outperforming VLP-mediated delivery (81.9%) at the highest dose. VLPs showed a dose-dependent decline in performance, likely due to cytotoxicity. In vivo, the deVINCI EVs achieved heart-selective delivery with robust DNA recombination in cardiac tissue and minimal off-target activity in peripheral organs, as confirmed by immunofluorescent analysis of tdTomato expression.
Conclusion
By integrating a modular architecture that enables high-capacity cargo loading and heart-selective delivery, the deVINCI technology establishes a scalable 'EV 2.0′ modular delivery platform for precision medicine.
| Original language | English |
|---|---|
| Article number | 57 |
| Journal | Cytotherapy |
| Volume | 28 |
| Issue number | 5 suppl. |
| DOIs | |
| Publication status | Published - May 2026 |
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