PhD Candidate Virginia Commonwealth University Richmond
This study focuses on optimizing mRNA lipid nanoparticles (LNPs) to enhance protein expression in macrophages, aiming for potential treatments of lung diseases. Using ethanol injection, we developed LNPs loaded with Firefly Luciferase mRNA, achieving high encapsulation efficiency (>75%), particle size ( < 300 nm), and low polydispersity index ( < 0.3). In vitro studies showed high transfection efficiency. We also achieved high transfection efficiency in RAW macrophage cells. A design of experiments (DOE) approach will be employed to optimize critical material attributes (CMAs) across lipid components; out of 4 factors each at 2 levels (Ionizable lipids:SM102 or ALC 315, phospholipids: DOPE, DPPC, PEG lipids: DMG-PEG2k, DSPE-PEG2k-Mannose, Sterol lipid: Cholesterol, β-sitosterol). To further enhance QTPP. Future work includes replacing Luciferase mRNA with therapeutic mRNA, stability testing in both fridge and freezing conditions, and aerosolization studies for nebulization applications.
Learning Objectives:
Describe key mRNA LNP components, demonstrate formulation optimization, understand transfection assays in macrophages and evaluate stability and aerosolization for therapeutic applications.
