In vivo mRNA delivery to virus-specific T cells by light-induced ligand exchange of MHC class I antigen-presenting nanoparticles

Abstract

Simultaneous delivery of mRNA to multiple populations of antigen (Ag)–specific CD8 + T cells is challenging given the diversity of peptide epitopes and polymorphism of class I major histocompatibility complexes (MHCI). We developed Ag-presenting nanoparticles (APNs) for mRNA delivery using pMHCI molecules that were refolded with photocleavable peptides to allow rapid ligand exchange by UV light and site-specifically conjugated with a lipid tail for postinsertion into preformed mRNA lipid nanoparticles. Across different TCR transgenic mouse models (P14, OT-1, and Pmel), UV-exchanged APNs bound and transfected their cognate Ag-specific CD8 + T cells equivalent to APNs produced using conventionally refolded pMHCI molecules. In mice infected with PR8 influenza, multiplexed delivery of UV-exchanged APNs against three immunodominant epitopes led to ~50% transfection of a VHH mRNA reporter in cognate Ag-specific CD8 + T cells. Our data show that UV-mediated peptide exchange can be used to rapidly produce APNs for mRNA delivery to multiple populations of Ag-specific T cells in vivo.

The landscape of mRNA nanomedicine

“…These EGFR-LNPs selectively delivered Cas9 mRNAs and single guide RNAs (sgRNAs) to disseminated EGFR-expressing ovarian tumors in mice, suppressing tumor growth and increasing the survival rate via efficient CRISPR-Cas9 gene editing. Selective delivery of mRNAs to antigen-specific CD8 + T cells 157 or CD4 + T cells 158 can be achieved by conjugating antigen or CD4 antibodies to LNPs, respectively. With more research, these organ-or cell-specific mRNA delivery platforms will expand the types of diseases that can be treated by mRNA therapies.…”

Optimal delivery strategies for nanoparticle-mediated mRNA delivery

“…61 Modifying LNPs with peptides is another method for targeting delivery of mRNA in vivo . Kwong et al 62 developed antigen (Ag)-presenting nanoparticles (APNs) for specific CD8+ T cells targeting by using UV-mediated peptide exchange which allowed the pMHC-I molecules to be post-inserted into the preformed mRNA LNPs. Those APNs showed successful mRNA delivery to multiple Ag-specific T cells in vivo .…”

“…60 Coupling exogenous target molecules on the surface of LNPs is a popular strategy to enhance delivery to specific cell populations or organs in vivo. The common exogenous target molecules include antibodies, 50,51,61 peptides, 62 natural ligands 63,64 and aptamers. 52 By using CD38 monoclonal antibody functionalized LNPs, a previous study constructed functional LNPs which specifically targeted to lymphoma cells.…”

Strategies for targeted gene delivery using lipid nanoparticles and cell-derived nanovesicles

“…When LNPs use ionizable cationic lipids, LNPs employing microfluidics and self-assembly processes outnumber LNPs employing thin film hydration and extrusion. 58–98 All the LNPs with permanently cationic lipids and nearly all the LNPs without cationic lipids use thin film hydration. 99–103 This implies the fact that the formulation method depends on the characteristics of the cationic lipids.…”