While lipid nanoparticles (LNPs) are widely used as efficient drug delivery systems in therapeutic modalities such as messenger ribonucleic acid (mRNA) vaccines, limitations in terms of targeting specificity continue to hinder their applications in precision medicine, particularly in the targeted delivery of drugs to specific retinal cell types, such as Müller cells. To enhance the targeting capability of LNPs toward Müller cells, the aim was to develop a novel LNP delivery system based on cell membrane (CM)-coating technology. Primary and immortalized Müller CMs were used to coat LNPs, and their targeting efficiency, transfection capability, and sustained action were systematically evaluated using in vitro co-culture assays and in vivo animal models. The LNPs coated with immortalized Müller CMs achieved significantly higher transfection efficiencies and better targeting for Müller cells compared to that of those coated with primary CMs; moreover, the former LNPs exhibited sustained drug release and targeting effects for up to three days, both in vitro and in vivo. Further proteomic analysis and functional validation of Achaete-scute homolog 1-loaded LNPs (LNP-Ascl1) and Müller CM-fused LNP-Ascl1 (CM-LNP-Ascl1) revealed that the latter likely promoted efficient Ascl1 delivery through vimentin (VIM)-mediated specific cellular recognition. This synergistic mechanism activated the Wnt/Janus kinase (JAK)-signal transducer and activator of transcription (STAT) signaling pathway and its downstream effector, matrix metalloproteinase 9 (MMP-9), establishing a positive feedback loop that facilitated cell proliferation. In summary, the immortalized Müller CM-coated LNPs outperformed their primary CM-coated counterparts in terms of both their targeting ability and sustained action. The stable membrane protein profile of immortalized membranes offers a promising strategy for refining precision drug delivery systems and demonstrates the potential for advancing targeted therapies for retinal diseases.
