Development of messenger RNA-loaded lipidoid-polymer hybrid nanoparticles
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Date
2021-07-07
Authors
López Cerdá, Sandra
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Abstract
Messenger RNA (mRNA) vaccines can be used for the treatment of non-infectious diseases and constitute a novel cancer immunotherapy modality. However, due to the challenges of mRNA delivery, the design of safe and efficient delivery systems is urgently needed. Lipidoid-polymer hybrid nanoparticles (LPNs), composed of a lipidoid and poly(DL-lactic-co-glycolic acid) (PLGA), are a novel type of nanocarriers that have been used for the delivery of small interfering RNA (siRNA) in the host lab. Nevertheless, siRNA and mRNA require distinct nanoparticle formulations. Thus, the aim of the present work was to explore the use of LPNs for mRNA delivery. mRNA-loaded LPNs modified with the C12-200 lipidoid were prepared by the double-emulsion solvent evaporation (DESE) method. C12-200 LPNs exhibited a z-average < 210 nm, a narrow size distribution and a nearly neutral zeta potential by dynamic light scattering. However, the mRNA appeared to be degraded during the probe sonication steps of the DESE method. Hence, microfluidics was harnessed for LPNs preparation to ensure mRNA integrity and allowed identification of the main bottleneck in the formulation design of C12-200 LPNs, which was the rigidity of the lipid shell. The combination of the L5N12 lipidoid with the helper lipid dioleylphosphatidylethanolamine (DOPE) in the lipid shell of LPNs resulted in an mRNA encapsulation efficiency of ~45% and an in vitro transfection efficiency > 80% in RAW 264.7 cells. A pilot study allowed optimization of relevant process and formulation parameters, i.e., a flow rate ratio of 10:1, a total flow rate of 12 mL/min and an L5N12:DOPE molar ratio of 3:1. However, the methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay revealed that L5N12-DOPE LPNs display 50% toxicity at the mRNA concentration that showed maximal transfection efficiency. Overall, this preliminary work paves the way to accelerate the design and development of an mRNA cancer vaccine using LPNs as delivery platform.
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Keywords
mRNA vaccines, Immunotherapy