Unlocking extracellular vesicles-mediated efficient DNA delivery for long-lasting transgene expression

Area of Science:

• Biotechnology
• Gene Therapy
• Extracellular Vesicles

Background:

• Gene therapy faces challenges in efficient DNA delivery for sustained transgene expression and minimizing immune responses.
• Red blood cell-derived extracellular vesicles (RBCEVs) are being explored as potential delivery vehicles.

Purpose of the Study:

• To characterize RBCEVs and evaluate their potential for DNA delivery in gene therapy.
• To achieve stable, prolonged transgene expression with reduced immune reactions.

Main Methods:

• Proteomic analysis of RBCEVs.
• Biodistribution studies in non-human primates and mice.
• In vivo delivery of luciferase and therapeutic plasmids (Factor IX, antibodies).
• Utilized cell-specific promoters for targeted expression.

Main Results:

• RBCEVs exhibit protein content similar to red blood cells, primarily hemoglobin.
• RBCEVs accumulate predominantly in the liver in non-human primates.
• Sustained luciferase expression for 13 months in mice.
• Durable Factor IX protein levels achieved.
• Simultaneous co-delivery of two plasmids demonstrated.
• Sustained antibody expression (Herceptin) achieved.
• Cell-specific promoters reduced immune responses.

Conclusions:

• RBCEVs represent a safe, scalable, and versatile platform for DNA delivery in gene therapy.
• RBCEVs enable sustained transgene expression and therapeutic protein production.
• RBCEV-mediated gene therapy holds potential for reduced treatment frequency and improved safety profiles.