Enhancing the efficacy and selectivity of lipid nanoparticles by optimizing their individual components

Abstract

This thesis investigates the rational design, optimization, and mechanistic characterization ofmRNA-LNP systems with emphasis on translational potency, intracellular delivery, tissueselectivetargeting, and structure–function relationships. The work integrates IVT mRNAengineering, lipid nanoparticle design, structural nanomedicine characterization, zebrafish-basedmechanistic screening, and imaging approaches to address current barriers in RNA nanomedicinedevelopment. Particular focus is placed on untranslated region optimization, the contribution ofKozak sequence variation to translation efficiency, the influence of lipid stereochemistry andphospholipid composition on LNP structure and function, and the mechanistic investigation ofnano-bio interactions governing biodistribution and intracellular delivery.Together, these studies address four interconnected dimensions of lipid nanoparticledevelopment: enhancing potency at the transcript level, elucidating LNP structure–functionrelationships, improving tissue- and cell-selective delivery, and extending lipid nanoparticledesign principles toward alternative therapeutic cargos. Across these areas, zebrafish embryosare employed as an in vivo platform to enable mechanistic investigation of LNP efficacy,biodistribution, tissue-selective targeting, and applications in cancer nanomedicine. Ultimately,this work aims to advance both mechanistic understanding and the rational engineering of nextgenerationLNP-based therapeutics.