Incorporation of cellular membrane protein extracts into lipid nanoparticles enhances their cellular uptake and mRNA delivery efficiency.

mRNA therapeutics enable transient expression of desired proteins within cells, holding great potential for advancements in vaccines, protein replacement therapies and gene editing approaches. Lipid nanoparticles (LNPs) are arguably the leading nanoplatform for mRNA delivery due to their scalability and transfection efficiency. However, their limited ability to target specific cell types, inefficient cellular uptake by many cell types, and endosomal entrapment represent challenges for improving targeted mRNA delivery. To address this, we evaluated a novel class of LNPs functionalized with cell-derived membrane proteins, that we refer to as hybrisomes. Membrane protein extracts (MPEs) were isolated from cultured cells using a mild detergent-based extraction protocol. Cy5-labeled mRNA encoding for eGFP was used to form LNPs and hybrisomes to investigate their internalization efficiency and mRNA delivery via flow cytometry and microscopy, with MPE content incorporated into hybrisomes during microfluidic mixing. MPEs were successfully incorporated into the lipid membrane of hybrisomes. Remarkably, the cellular uptake of hybrisomes was up to 15-fold higher than LNPs, while the mRNA delivery efficiency improved up to 8-fold depending on the MPE content incorporated into the hybrisomes. Further studies confirmed that the enhanced cellular uptake of hybrisomes and mRNA is partially explained by the presence of membrane proteins and hybrisomes' unique morphology including bleb-like structures. Moreover, the versatility of hybrisomes was demonstrated by producing formulations using MPEs isolated from different cell types, which led to variations in cellular uptake and mRNA delivery, suggesting that the cell type from which MPEs are derived influences their biological function. These findings pave the way for the development of more targeted and effective nanotherapeutic strategies.