Lipid-peptide nanocomplexes for mRNA delivery in vitro and in vivo

Dania Grant-Serroukh; Morag R Hunter; Ruhina Maeshima; Aristides D Tagalakis; Ahmad M Aldossary; Nour Allahham; Gareth R Williams; Mark Edbrooke; Arpan Desai; Stephen L Hart

doi:10.1016/j.jconrel.2022.06.018

Lipid-peptide nanocomplexes for mRNA delivery in vitro and in vivo

J Control Release. 2022 Aug:348:786-797. doi: 10.1016/j.jconrel.2022.06.018. Epub 2022 Jun 26.

Affiliations

¹ UCL Great Ormond Street Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, UK; UCL School of Pharmacy, University College London, 29 - 39 Brunswick Square, London WC1N 1AX, UK.
² Pharmaceutical Sciences, AstraZeneca, Aaron Klug Building, Granta Park, Cambridge CB21 6GH, UK.
³ UCL Great Ormond Street Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, UK.
⁴ UCL School of Pharmacy, University College London, 29 - 39 Brunswick Square, London WC1N 1AX, UK.
⁵ UCL Great Ormond Street Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, UK. Electronic address: s.hart@ucl.ac.uk.

PMID: 35718210
DOI: 10.1016/j.jconrel.2022.06.018

Abstract

Despite recent advances in the field of mRNA therapy, the lack of safe and efficacious delivery vehicles with pharmaceutically developable properties remains a major limitation. Here, we describe the systematic optimisation of lipid-peptide nanocomplexes for the delivery of mRNA in two murine cancer cell types, B16-F10 melanoma and CT26 colon carcinoma as well as NCI-H358 human lung bronchoalveolar cells. Different combinations of lipids and peptides were screened from an original lipid-peptide nanocomplex formulation for improved luciferase mRNA transfection in vitro by a multi-factorial screening approach. This led to the identification of key structural elements within the nanocomplex associated with substantial improvements in mRNA transfection efficiency included alkyl tail length of the cationic lipid, the fusogenic phospholipid, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and cholesterol. The peptide component (K₁₆GACYGLPHKFCG) was further improved by the inclusion of a linker, RVRR, that is cleavable by the endosomal enzymes cathepsin B and furin, and a hydrophobic motif (X-S-X) between the mRNA packaging (K₁₆) and receptor targeting domains (CYGLPHKFCG). Nanocomplex transfections of a murine B16-F10 melanoma tumour supported the inclusion of cholesterol for optimal transfection in vivo as well as in vitro. In vitro transfections were also performed with mRNA encoding interleukin-15 as a potential immunotherapy agent and again, the optimised formulation with the key structural elements demonstrated significantly higher expression than the original formulation. Physicochemical characterisation of the nanocomplexes over time indicated that the optimal formulation retained biophysical properties such as size, charge and mRNA complexation efficiency for 14 days upon storage at 4 °C without the need for additional stabilising agents. In summary, we have developed an efficacious lipid-peptide nanocomplex with promising pharmaceutical development properties for the delivery of therapeutic mRNA.

Keywords: Lipids; Nanocomplexes; Peptides; Targeting; Transfection; mRNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Humans
Lipids / chemistry
Liposomes* / chemistry
Melanoma*
Mice
Peptides / chemistry
RNA, Messenger / genetics
Transfection

Substances

Lipids
Liposomes
Peptides
RNA, Messenger