Evaluation of High-throughput Extracellular Vesicle Loading Platform for Therapeutic Genome Editing INDs

ABSTRACT Extracellular vesicles (EVs) have been an emerging approach for delivering genome editing therapeutics. Due to their superior biocompatibility and low immunogenicity compared to lipid nanoparticles (LNPs) and viral vectors, EVs have been in increased needs for developing gene therapy. Despite great promise for clinical translation, standardization of EV isolation and cargo loading at the scale has been challenging, which substantially hindered the clinical translation of EV-based genome editing technologies. Transfection of EVs for genome editor loading is a key step in functional therapeutic development. However, current transfection approaches suffer from low transfection rate, and low scalability and volume processing throughput. In this project, we aim to optimize and evaluate our recently developed high-throughput EV loading platform with rigorous quality characterization on EVs and genome editor payload, which will provide essential data on CMC, Critical Quality Attributes (CQAs), and toxicology of EV based CRISPR gene therapy for IND submission. The Aim 1 will focus on optimization of high-throughput EV loading platform (µDES) with genome editors for GMP manufacturing. The Aim 2 will optimize ExoQuality approach as the IND-enabling assessment to establish the Critical Quality Attributes (CQAs) of EV based genome editing therapeutic products. The Aim 3 will focus on the pharmacological and toxicological evaluation on developed CRISPR RNP EVs with modality in treating hearing loss for IND submission. The project will lead to the scalable GMP manufacturing procedures for EV based genome editing products and establish acceptable CQA criteria and CMC data to accelerate EV based genome editing product for IND submissions. The outcome will significantly impact on accelerating the translation of novel, safe, and effective therapeutic genome editing strategies to first-in-human clinical trials.