Ionizable Lipids Drive Subcellular Localization and Immune Cell Targeting of Barcoded Nanoparticles in Lung Cancer

Area of Science:

• Cellular Biology
• Nanotechnology
• Biomedical Imaging

Background:

• Drug delivery efficacy depends on nanoparticle (NP) interaction with target cells and subcellular organelles.
• Organelle dysfunction is implicated in numerous diseases, highlighting the need to track NP destinations.
• Existing methods lack precision in quantifying multiple NP types and their intracellular localization.

Purpose of the Study:

• To develop and validate a novel microscopy technique for simultaneous quantification of multiple nanoparticle types.
• To precisely analyze the intracellular and inter-cellular distribution of core-shell nanoparticles (NPs) in vitro and in vivo.
• To investigate the subcellular organelle destinations of nucleic acids delivered by different cationic lipids.

Main Methods:

• Nanoparticle (NP) microscopy utilizing signal amplification of DNA barcodes.
• Multiplexed cyclic immunofluorescence technique for simultaneous NP quantification.
• In vitro and in vivo studies using three core-shell NPs (G0-P5, 7C1-F5, and C12-D).

Main Results:

• The developed technique enhanced signal-to-noise ratio by 15-fold compared to standard fluorescence in situ hybridization.
• In vitro: Nucleic acids with G0-C14 cationic lipids localized in lysosomes (macrophages) and mitochondria (tumor cells).
• In vivo: G0-C14 cationic lipids showed highest uptake by CD206+ immune cells; 7C1 and C12-200 lipids were preferentially taken up by CD206+CD11c+Arg1+ immune cells.

Conclusions:

• The novel NP microscopy technique enables precise, simultaneous quantification of multiple NP types and their subcellular localization.
• Understanding NP organelle destinations is crucial for predicting drug effects and enhancing potency.
• The findings provide insights into targeted delivery strategies for immune cells and therapeutic applications.