Impact of Ionizable Lipid Variation on the Immunogenicity of Lipid Nanoparticles

Authors

Chloe M. Krueger, DePauw University
Abbey L. Stokes, University of Arkansas
Shilpi Agrawl, University of Arkansas
Christopher E. Nelson, University of Arkansas

Files

Document Type

Poster

Publication Date

10-1-2025

Abstract

Lipid nanoparticles (LNPs) are a leading nonviral delivery system for nucleic acid therapeutics due to their scalability and efficiency. However, certain formulations may trigger undesired immune responses. This study aimed to assess the inflammatory potential of LNPs formulated with different ionizable lipids using a murine macrophage reporter cell line (RAW-IRCs). RAW-IRCs express GFP upon successful mRNA delivery and mRFP1 upon activation of inflammatory pathways. A library of LNPs was synthesized via vortex mixing and characterized for hydrodynamic diameter, polydispersity index, and encapsulation efficiency. Treated RAW-IRCs were analyzed by flow cytometry to evaluate GFP and mRFP1 expression. Our results highlight seven ionizable lipids that show higher transfection with a lower inflammatory response. Additionally, when RAW-IRCs were treated with lipopolysaccharide (LPS), an inflammatory stimulant, there is a drop in transfection efficiency regardless of the ionizable lipid being changed. These findings contribute to the rational design of LNPs with reduced immunogenicity for safer gene therapy applications.

Department

Department of Biomedical Engineering, University of Arkansas, Fayetteville, ARDepartment of Chemistry & Biochemistry, DePauw University, Greencastle, INDepartment of Cell and Molecular Biology, University of Arkansas, Fayetteville, AR

Project Mentor

Christopher E. Nelson

Funding and Acknowledgements

CK was supported by the NSF REU Program (EEC 2243953 PI: Dr. Muldoon) ALS was supported by the Distinguished Doctoral Fellowship Dr. Nelson's lab is funded by NIH R35GM155433 Work supported by Women’s Giving Circle Dr. Kim Lab for the use of their DLS Machine Dr. Samsonraj’s Lab for the use of their plate reader

Recommended Citation

Krueger, Chloe M.; Stokes, Abbey L.; Agrawl, Shilpi; and Nelson, Christopher E., "Impact of Ionizable Lipid Variation on the Immunogenicity of Lipid Nanoparticles" (2025). Annual Student Research Poster Session. 203.
https://scholarship.depauw.edu/srfposters/203