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

• DOI: 10.1016/j.jconrel.2022.06.018
• Publication Date: 2022-06-26T08:52:52Z
• Authors: Dania Grant-Serroukh; Morag R. Hunter; Ruhina Maeshima; Aristides D. Taga...; Ahmad M. Aldossary; Nour Allahham; Gareth R. Williams; Mark Edbrooke; Arpan Desai; Stephen L. Hart
• Keywords: Targeting; 0303 health sciences; mRNA; Transfection; Lipids; 3. Good health; Mice; 03 medical and health sciences; Liposomes; Animals; Humans; Nanocomplexes; RNA, Messenger; Peptides; Melanoma

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