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Structural Polymorphism of Solid Lipid Nanoparticles (LNPs) containing Natural Cholesterol Analogs


EMSL Project ID
51108

Abstract

Lipid-based nanoparticles (LNPs) are the most advanced nanotherapeutics for the delivery of ribonucleic acids (RNA) for gene therapy. They are the only nanoparticles being used in the clinic for RNA delivery. Yet, there is currently a lack of understanding of the relationship between LNP morphology and their therapeutic potential. Recently, our research group investigated the incorporation of natural cholesterol analogs (namely, ?-sitosterol, stigmastanol, campesterol, and fucosterol; resulting particles further referred to as enhanced LNPs or eLNPs) in LNP formulation and found significant (up to 50-fold) improvements in gene delivery. Preliminary cryo-TEM micrographs indicate that ?-sitosterol eLNPs have a highly faceted surface, as opposed to spherical cholesterol LNPs. We propose conducting a series of cryotomography experiments on LNPs containing cholesterol and its analogs to deepen the understanding of the particle morphology and its influence on the efficiency of gene delivery. The specific aims of this project are: 1) receive initial training on sample optimization, data collection and processing; 2) collect, process, and interpret cryotomography data for LNPs and ?-sitosterol eLNPs; and 3) optimize sample preparation and acquire preliminary images for stigmastanol, campesterol, and fucosterol eLNPs to determine appropriateness of future studies for comparative analysis using cryo-ET. If successful, the findings of this study will provide unique insights into the correlation between eLNP morphology and the potency of cargo delivery, thus helping to establish eLNP design guidelines for next-generation gene therapeutics.


Project Details

Start Date
2019-10-15
End Date
2020-08-15
Status
Closed

Team

Principal Investigator

Gaurav Sahay
Institution
Oregon State University

Team Members

Siddharth Patel
Institution
Oregon State University

Yulia Eygeris
Institution
Oregon State University

Theo Humphreys
Institution
Oregon Health & Science University

Related Publications

Y. Eygeris, S. Patel, A. Jozic, G. Sahay, Deconvoluting Lipid Nanoparticle Structure for Messenger RNA Delivery Nano Lett. 2020, 20, 4543?4549.