Repurposing an endogenous human mRNA transfer system as a gene therapy tool

This project aims to map out the feasibility and performance of a naturally-ocurring system for mRNA transfer in humans when repurposed to deliver recombinant RNA payloads. The system involves a single protein that, upon translation, associates with its encoding mRNA then forms a particle that sequesters that mRNA, after which the particle is enveloped within vesicles that bud into a multivesicular body (MVB) before cellular release and virus-like transfer to target cells in which the payload mRNA is released for subsequent translation.

Gene therapy approaches have recently faced challenges due in part to adverse effects arising from the immunogenicity of the viral vectors used for tgene transfer. Kumin et al. (2021) report that, on average, 35% of 149 2021 AAV gene therapy clinical trials were associated with treatment-emergent serious adverse events.

In this project the Protein X mRNA will be modified to test its capacity to incorporate recombinant RNA elements. Inserted elements of differing size and sequence will be tested for their ability to be transferred from packaging cells to target cells via the engineerred Protein X system. It is anticipated that the immunogenicity of a Protein-X system will be significantly lower than that of virus-based systems.