Rerouting mRNA for in vivo chimeric antigen receptor immune cell therapy

Chimeric antigen receptor (CAR) T cell therapy has revolutionized cancer treatment. Nevertheless, CAR T cell therapy is clinically limited to hematological malignancies and hampered by its complex and costly ex vivo manufacturing procedure. To render this treatment cost-effective and available for diverse cancer types, in vivo CAR immune cell therapy strategies must be developed. My group has established new apolipoprotein nanoparticle (aNP) platform technology for RNA delivery to immune cells. Leveraging lipoproteins’ natural delivery features, our technology combines tunable biodistribution features with a superb biocompatibility profile. Building on this concept, I propose developing aNPs containing messenger RNA (mRNA) encoding CARs to be delivered and expressed by specific immune cells.Based on compelling preliminary data, I will pursue two Key Objectives:KO1: Using advanced microfluidic production methods, we will establish a library of self-assembling aNPs – composed of natural phospho(lipids), ionizable cationic materials and apolipoprotein A1 (apoA1), and containing mRNA – and screen barcoded aNPs’ ability to functionally deliver mRNA to immune cells in vivo. Machine learning analysis will provide lead formulations for in vivo CAR monocyte/macrophage (CAR M) proof-of-concept therapeutic studies in a solid tumor mouse model.KO2: Using proprietary apoA1-nanobody fusion protein technology, we will screen targeted mRNA-aNPs’ ability to induce functional gene expression in specific immune cells in vivo. Lead aNP-mRNA formulations will be used for in vivo CAR T proof-of-concept therapeutic studies in a hematological mouse tumor model.This Program’s successful completion will yield a unique, versatile nanomedicine approach that can be employed to induce in vivo CAR immune cell therapy for cancer treatment, with significant translational potential and pertinency for applications in non-malignant diseases such as autoimmune disorders and cardiovascular disease.