biostudies-literatureHan ELNational Center for Advancing Translational SciencesAmerican Cancer SocietyNCATS NIH HHSNIDCR NIH HHSNHLBI Division of Intramural ResearchNHLBI NIH HHSNational Institute of Dental and Craniofacial ResearchDivision of Chemical, Bioengineering, Environmental, and Transport SystemsDivision of Graduate Education1477-1486https://www.ebi.ac.uk/biostudies/studies/S-EPMC11922166biostudies-literatureUnknown24(5)Lipid nanoparticle (LNP)-mediated nucleic acid therapies, including mRNA protein replacement and gene editing therapies, hold great potential in treating neurological disorders including neurodegeneration, brain cancer, and stroke. However, delivering LNPs across the blood-brain barrier (BBB) after systemic administration remains underexplored. In this work, we engineered a high-throughput screening transwell platform for the BBB (HTS-BBB), specifically optimized for screening mRNA LNPs. Unlike most transwell assays, which only assess transport across an endothelial monolayer, HTS-BBB simultaneously measures LNP transport and mRNA transfection of the endothelial cells themselves. We then use HTS-BBB to screen a library of 14 LNPs made with structurally diverse ionizable lipids and demonstrNano lettersPredictive High-Throughput Platform for Dual Screening of mRNA Lipid Nanoparticle Blood-Brain Barrier Transfection and Crossing.PMC11922166F30HL16246501A1T90DE030854F30 HL162465T90 DE030854RSG-22-122-01-ETDP2 TR002776CBET-21454911845298Kim DMitchell MJHan ELPadilla MSPalanki RMrksich KYoon ICTang SLi JJfalsePredictive High-Throughput Platform for Dual Screening of mRNA Lipid Nanoparticle Blood-Brain Barrier Transfection and Crossing.Lipid nanoparticle (LNP)-mediated nucleic acid therapies, including mRNA protein replacement and gene editing therapies, hold great potential in treating neurological disorders including neurodegeneration, brain cancer, and stroke. However, delivering LNPs across the blood-brain barrier (BBB) after systemic administration remains underexplored. In this work, we engineered a high-throughput screening transwell platform for the BBB (HTS-BBB), specifically optimized for screening mRNA LNPs. Unlike most transwell assays, which only assess transport across an endothelial monolayer, HTS-BBB simultaneously measures LNP transport and mRNA transfection of the endothelial cells themselves. We then use HTS-BBB to screen a library of 14 LNPs made with structurally diverse ionizable lipids and demonstr2024-01-01T00:00:00Z2024 Feb2025-04-04T14:47:39.2Z2025-04-04T14:47:39.2ZS-EPMC119221663825919810.1021/acs.nanolett.3c03509