The development of a targeted delivery system for saRNAs for immunotherapy of monocyte-derived leukemia

Immunotoxins (ITs) are representing a subset of antibody drug conjugates where the synthetic toxin is replaced by a bacterial protein toxin. Major drawback is related to the immunogenicity of the protein not allowing repeated application. Consequently targeted human cytolytic fusion proteins (hCFPs) have been developed to replace the foreign protein toxin by a human apoptosis inducing enzyme exemplified by human angiogenin (hAng). Currently such hCFPs show reduced cytotoxicity compared to ITs, as it depends on efficient delivery of sufficient amounts into the cytosol. Therefore, this collaboration research planned to develop a targeted saRNA delivery system to introduce hAng encoding saRNA into the cytosol of an antigen-positive target cell. saRNA produces copies of itself which should increase the amount of enzyme to effectively kill a cancer cell.

Methods: eGFP/hAng encoding saRNA sequence will be cloned into a plasmid. The linearized vector will be transcribed and purified using LiCl. Purified saRNA will be encapsulated in a lipid nanoparticle decorated with CD64-specific antibody fragments. The LNP-eGFP-saRNA conjugates will be delivered into CD64-positive leukemia cell lines. Once proof of concept is confirmed the saRNA encoding eGFP will be replaced by saRNA encoding hAng for selective elimination of monocyte-derived leukemia cells.

Faculty Supervisor:

Anna Blakney

Student:

Partner:

University of Cape Town

Discipline:

Life Sciences

Sector:

Biotechnology; Health and Related Sciences & Technology; Nanotechnology

University:

The University of British Columbia

Program: