Leiden University Scholarly Publications

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Escalona Rayo, O. (2026)

Lipid nanoparticle technology for mRNA delivery: bridging vaccine applications with fundamental insights into nano-bio interactions

Doctoral Thesis
Lipid nanoparticles (LNPs) have enabled the success of mRNA vaccines but remain limited in broader therapeutic use by challenges in delivery efficiency, targeting, and mechanistic understanding. This thesis aimed to advance the rational design of mRNA–LNP systems by systematically examining how lipid composition governs delivery performance, immune activation, and nano–bio interactions across in vitro and in vivo models. Comparative analysis of ionizable lipids showed that while LNPs shared similar physicochemical properties and mRNA encapsulation efficiency, their biological activity was highly context dependent, with in vitro potency poorly predicting in vivo protein expression and immune responses. To overcome the weak immunogenicity of tumor antigens, a heterologous prime–boost vaccination strategy combining antigen-encoding mRNA-LNPs with costimulatory agonist-based boosters was developed, significantly enhancing the magnitude and durability of antigen...Show moreLipid nanoparticles (LNPs) have enabled the success of mRNA vaccines but remain limited in broader therapeutic use by challenges in delivery efficiency, targeting, and mechanistic understanding. This thesis aimed to advance the rational design of mRNA–LNP systems by systematically examining how lipid composition governs delivery performance, immune activation, and nano–bio interactions across in vitro and in vivo models. Comparative analysis of ionizable lipids showed that while LNPs shared similar physicochemical properties and mRNA encapsulation efficiency, their biological activity was highly context dependent, with in vitro potency poorly predicting in vivo protein expression and immune responses. To overcome the weak immunogenicity of tumor antigens, a heterologous prime–boost vaccination strategy combining antigen-encoding mRNA-LNPs with costimulatory agonist-based boosters was developed, significantly enhancing the magnitude and durability of antigen-specific T cell responses and revealing lipid-dependent, tissue-specific immune effects. Further investigation into nano–bio interactions highlighted the role of helper lipid charge in macrophage targeting, demonstrating that cationic helper lipids improve endosomal escape and mRNA delivery but raise safety considerations. Finally, chemically modified, clickable ionizable lipids combined with advanced imaging approaches enabled super-resolution visualization of LNP intracellular trafficking and disassembly, providing powerful tools to elucidate mechanisms underlying mRNA-LNP delivery.Show less
mRNA delivery
Lipid nanoparticles
Vaccines
Nano-bio interactions
Clickable ionizable lipids
All authors
Escalona Rayo, O.
Supervisor
Kros, A.; Jiskoot, W.; Slütter, B.A.
Committee
Ubbink, M.; Bouwstra, J.A.; Schneider, G.F.; Meel, R. van der; Jakopin, Z.
Qualification
Doctor (dr.)
Awarding Institution
Leiden Institute of Chemistry (LIC), Faculty of Science, Leiden University
Date
2026-01-20

Funding

Sponsorship
SECIHTI-Mexico [CVU/grant number: 619711/766609]