Ly, H.H., Daniel, S., Soriano, S.K.V. et al. (2 more authors) (2022) Optimization of lipid nanoparticles for saRNA expression and cellular activation using a design-of-experiment approach. Molecular Pharmaceutics, 19 (6). pp. 1892-1905. ISSN 1543-8384
Abstract
Lipid nanoparticles (LNPs) are the leading technology for RNA delivery, given the success of the Pfizer/BioNTech and Moderna COVID-19 mRNA (mRNA) vaccines, and small interfering RNA (siRNA) therapies (patisiran). However, optimization of LNP process parameters and compositions for larger RNA payloads such as self-amplifying RNA (saRNA), which can have complex secondary structures, have not been carried out. Furthermore, the interactions between process parameters, critical quality attributes (CQAs), and function, such as protein expression and cellular activation, are not well understood. Here, we used two iterations of design of experiments (DoE) (definitive screening design and Box-Behnken design) to optimize saRNA formulations using the leading, FDA-approved ionizable lipids (MC3, ALC-0315, and SM-102). We observed that PEG is required to preserve the CQAs and that saRNA is more challenging to encapsulate and preserve than mRNA. We identified three formulations to minimize cellular activation, maximize cellular activation, or meet a CQA profile while maximizing protein expression. The significant parameters and design of the response surface modeling and multiple response optimization may be useful for designing formulations for a range of applications, such as vaccines or protein replacement therapies, for larger RNA cargoes.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2022 The Authors. Published by American Chemical Society. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives licence (https://creativecommons.org/licenses/by-nc-nd/4.0/). |
Keywords: | Box−Behnken Design; cytokine response; definitive screening design; design-of-experiment (DoE); lipid nanoparticle (LNP); mRNA (mRNA); protein expression; self-amplifying mRNA (saRNA); Amino Alcohols; COVID-19; Caprylates; Decanoates; Humans; Liposomes; Nanoparticles; RNA, Messenger; RNA, Small Interfering |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Engineering (Sheffield) > Department of Chemical and Biological Engineering (Sheffield) |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 12 Jan 2024 11:55 |
Last Modified: | 12 Jan 2024 11:55 |
Published Version: | http://dx.doi.org/10.1021/acs.molpharmaceut.2c0003... |
Status: | Published |
Publisher: | American Chemical Society (ACS) |
Refereed: | Yes |
Identification Number: | 10.1021/acs.molpharmaceut.2c00032 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:207529 |