Abstract

Extracellular Vesicle-Based Gene Therapy

Mei He, Associate Professor, University of Florida

Genome editing is an emerging and powerful therapeutic tool for treating diverse diseases. However, clinical translation has been challenging, due to tremendous limitations in current delivery vehicles such as traditional viral vectors for carrying CRISPR Cas9 systems. Alternatively, extracellular vesicles (EV) based gene delivery is emerging as a safe and highly biocompatible approach for addressing current challenges in gene therapy. We developed a novel Microfluidic Droplet-based EV Electroporation System (µDES), which can handle various cargos loaded into EVs in large throughput and high efficiency. We achieved 10-fold enhancement of loading efficiency and more than 1000-fold increase in processing throughput on loading CRISPR RNP complexes into EVs compared with conventional bulk electroporation. In the Shaker-1 mouse model of dominant progressive hearing loss, we demonstrated the effective delivery of RNP-EVs into inner ear hair cells, with a clear reduction of Myo7ash1 mRNA expression compared to RNP-loaded lipid-like nanoparticles (RNP-LNPs), leading to significant hearing recovery for future clincial translation.