In vivo imaging of lipid nanoparticle messenger RNA therapeutics: Current advances and emerging opportunities.

Non-viral and non-lipid nanoparticles for RNA therapeutics: Design, applications, and preclinical studies.

This review examines non-viral and non-lipid RNA nanocarriers from mechanistic and translational perspectives and highlights non-viral and non-lipid nanocarriers as complementary platforms for context-specific RNA therapy.

Targeted delivery of mRNA to immune cells for in vivo cell therapy

The integration of rational material design, high-throughput screening, artificial intelligence, and interdisciplinary collaboration will be essential to advance next-generation targeted in vivo mRNA cell therapies toward clinical translation.

Lipid–ASO therapeutics exhibit differential tissue targeted delivery upon systemic or local CNS administration

Nitric oxide-assisted lipid nanoparticles amplify mRNA vaccine responses.

mRNA vaccines have made substantial clinical advances, yet their full clinical potential can be further expanded by enhancing cytosolic delivery. Here, we integrate a nitric oxide (NO) generator with lipid nanoparticles (LNPs) to boost mRNA delivery efficiency and mRNA-based vaccine efficacy. SM-102/DEA LNPs, the lead formulation, achieved significantly higher mRNA delivery compared with the FDA approved SM-102 LNPs in both cellular and animal models. The intramuscular administration of SM-102/DEA LNPs encapsulating mRNA encoding SARS-CoV-2 spike protein elicited substantially higher anti-spike IgG levels and robust CD8+ and CD4+ T cell responses compared to SM-102 LNPs. Mechanistic studies revealed that DEA incorporation promotes endosomal escape of mRNA cargos in SM-102/DEA LNPs. These findings establish NO-assisted LNPs as a unique platform for potent mRNA delivery, which provides a new paradigm for overcoming endosomal barriers and improving the efficacy of mRNA vaccines.

Construction of a translational whole-body physiologically based pharmacokinetic model to investigate the biodistribution of lipid nanoparticle-based mRNA therapeutics for intracellular protein targets

Lipid nanoparticle (LNP)–based messenger RNA (mRNA) therapeutics enable in vivo production of diverse protein modalities, but their translation is complicated by multiscale pharmacokinetics encompassing tissue transport, cellular uptake, and intracellular processing. We developed a platform translational whole-body physiologically based pharmacokinetic (PBPK) model to mechanistically characterize the biodistribution and protein expression of intravenously administered mRNA–LNP therapeutics. The model integrates receptor-mediated, saturable tissue infiltration, lymphatic recirculation, hepatic uptake, and explicit intracellular processes including endosomal degradation, mRNA escape, translation, and protein turnover. Parameters were calibrated using comprehensive rat biodistribution data containing both tissue-level mRNA and translated protein measurements, enabling characterization of rapid early tissue deposition and dominant hepatic protein production. Sensitivity analyses identified endosomal degradation, mRNA stability, and translation efficiency as the primary determinants of hepatic protein exposure, whereas increasing tissue influx alone yielded minimal effect. Using fundamental allometric scaling principle, the model was translated from rat to human and evaluated against published clinical plasma mRNA pharmacokinetics of an mRNA-encoded monoclonal antibody. Virtual population simulations reproduced dose-dependent peak timing and clearance across multiple dosing regimens. Cell-type specific simulation analyses further demonstrated the competing roles of Kupffer cells and hepatocytes in shaping liver protein exposure. This whole-body PBPK framework provides a mechanistic and predictive platform to support first-in-human dose selection, to interrogate multilevel determinants of mRNA pharmacokinetics, and to guide rational mRNA-LNP design.

Targeted mRNA Delivery Using Bispecific Antibody-Lipid Nanoparticle Complexes.

Lipid nanoparticles (LNPs) are highly modular drug nanocarriers that have significantly progressed in the development and application of novel RNA therapeutics. While instrumental in current vaccine and hepatic gene therapy applications, extrahepatic LNP delivery remains a challenging hurdle, severely limiting clinical applications for individualized therapies that require cellular- and tissue-specific enrichment. Novel platform development remains needed to generate targeted LNPs capable of modeling targeted uptake in vitro prior to in vivo applications. Herein, we developed and utilized an antibody-based targeting bispecific-LNP (TbsAb-LNP) platform simultaneously binding LNP polyethylene glycol motifs and desired cell surface markers. By colocalizing TbsAb-LNP complexes with desired cell surface markers, we enable mRNA-LNP accumulation in the desired cell types. We additionally assessed the endosomal efficiency kinetics of TbsAb-LNP complexes generated at various bispecific loading ratios in a Galectin-9 in vitro expression model. Leveraging this facile and modular platform that eliminates the need for complex chemical conjugation approaches, we successfully achieved targeted LNP delivery to xhuCD3-expressing lymphoblasts and xmTfR1-expressing cell lines in vitro as well as xmTfR1-expressing cells in vivo. Enrichment of TbsAb-LNPs to splenic cells demonstrates the potential utility of this platform for clinical vaccine and T-cell delivery applications.