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Correlating the Structure and Gene Silencing Activity of Oligonucleotide-Loaded Lipid Nanoparticles Using Small-Angle X-ray Scattering.
| Content Provider | Europe PMC |
|---|---|
| Author | Hammel, Michal Fan, Yuchen Sarode, Apoorva Byrnes, Amy E. Zang, Nanzhi Kou, Ponien Nagapudi, Karthik Leung, Dennis Hoogenraad, Casper C. Chen, Tao Yen, Chun-Wan Hura, Greg L. |
| Copyright Year | 2023 |
| Abstract | With three FDA-approvedproducts, lipid nanoparticles (LNPs) areunder intensive development for delivering wide-ranging nucleic acidtherapeutics. A significant challenge for LNP development is insufficientunderstanding of structure–activity relationship (SAR). Smallchanges in chemical composition and process parameters can affectLNP structure, significantly impacting performance in vitro and in vivo. The choice of polyethylene glycollipid (PEG-lipid), one of the essential lipids for LNP, has been provento govern particle size. Here we find that PEG-lipids can furthermodify the core organization of antisense oligonucleotide (ASO)-loadedLNPs to govern its gene silencing activity. Furthermore, we also havefound that the extent of compartmentalization, measured by the ratioof disordered vs ordered inverted hexagonal phases within an ASO-lipidcore, is predictive of in vitro gene silencing. Inthis work, we propose that a lower ratio of disordered/ordered corephases correlates with stronger gene knockdown efficacy. To establishthese findings, we developed a seamless high-throughput screeningapproach that integrated an automated LNP formulation system withstructural analysis by small-angle X-ray scattering (SAXS) and in vitro TMEM106b mRNA knockdown assessment. We appliedthis approach to screen 54 ASO-LNP formulations while varying thetype and concentration of PEG-lipids. Representative formulationswith diverse SAXS profiles were further visualized using cryogenicelectron microscopy (cryo-EM) to help structural elucidation. Theproposed SAR was built by combining this structural analysis with in vitro data. Our integrated methods, analysis, and resultingfindings on PEG-lipid can be applied to rapidly optimize other LNPformulations in a complex design space. |
| ISSN | 19360851 |
| Journal | ACS Nano |
| Volume Number | 17 |
| PubMed Central reference number | PMC10311593 |
| Issue Number | 12 |
| PubMed reference number | 37279108 |
| e-ISSN | 1936086X |
| DOI | 10.1021/acsnano.3c01186 |
| Language | English |
| Publisher | American Chemical Society |
| Publisher Date | 2023-06-06 |
| Access Restriction | Open |
| Rights License | Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/). © 2023 The Authors. Published by American Chemical Society |
| Subject Keyword | lipid nanoparticle small-angle X-ray scattering cryogenic electron microscopy high-throughput screening PEG-lipid structure−activity relationship |
| Content Type | Text |
| Resource Type | Article |
| Subject | Nanoscience and Nanotechnology Physics and Astronomy Engineering Materials Science |
