AI-Validated Brain Targeted mRNA Lipid Nanoparticles with Neuronal Tropism

Targeting therapeutic nanoparticles to the brain poses a challenge due to the restrictive nature of the blood-brain barrier (BBB). Here we report the development of mRNA-loaded lipid nanoparticles (LNPs) functionalized with BBB-interacting small molecules, thereby enhancing brain delivery and gene expression. Screening brain-targeted mRNA-LNPs in central nervous system (CNS) <i>in vitro</i> models and through intravenous administration in mice demonstrated that acetylcholine-conjugated LNPs achieved superior brain tropism and gene expression, outperforming LNP modifications with nicotine, glucose, memantine, cocaine, tryptophan, and other small molecules. An artificial intelligence (AI)-based model designed to predict the BBB permeability of small-molecule ligands showed strong alignment with our experimental results, providing <i>in vivo</i> validation of its predictive capacity. Cell-specific biodistribution analysis in Cre-reporter Ai9 mice showed that acetylcholine-functionalized LNPs preferentially transfected neurons and astrocytes following either intravenous or intracerebral administration. Mechanistic studies suggest that acetylcholine-LNP uptake is mediated by the functional engagement of acetylcholine receptors (AchRs) followed by endocytosis, which synergistically enhances intracellular mRNA delivery. Moreover, acetylcholine-LNPs successfully crossed a human BBB-on-a-chip model, enabling transgene expression in human iPSC-derived neurons. Their effective penetration and transfection in human brain organoids further support their potential activity in human-based systems. These findings establish a predictive and modular framework for engineering CNS-targeted LNPs, advancing precision gene delivery for brain disorders.

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Publication Details

Journal
ACS Nano
Published
2025-09-16
DOI
https://doi.org/10.1021/acsnano.4c15013
Citations
18
Primary Topic
ATP Synthase and ATPases Research
Type
article
Field-Weighted Citation Impact
14.51
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article

AI-Validated Brain Targeted mRNA Lipid Nanoparticles with Neuronal Tropism

Inbar Freilich, Avi Schroeder, Galoz Kaneti, Tal Laviv et al.
18 citations
ACS Nano
ATP Synthase and ATPases Research
14.51
article

AI-Validated Brain Targeted mRNA Lipid Nanoparticles with Neuronal Tropism

Inbar Freilich, Avi Schroeder, Galoz Kaneti, Tal Laviv, Gal Chen, Uri Ashery, Egor Egorov, Yehuda Arav, Amit Zeisel, Mor Sela, Raneen Nicola, Kira Radinsky, Shaked Kagan, Ben M. Maoz, Keshet Tadmor, Tommaso Patriarchi, Thomas A. Schroeder, Inbal Abutbul‐Ionita, Peleg Hasson, Tomer Kagan, Yuval Richtman, Jeny Shklover, Yael Leichtmann‐Bardoogo, Ibrahim Knani, Dganit Danino, Patricia Mora‐Raimundo, Sally Turutov, Anas Odeh, Reaam Kablan, Tasneem Abu-Raiya, Haim Kadosh, Lin Zhige, Mia R. Albalak Menasherov
article en
18 citations

Abstract

Targeting therapeutic nanoparticles to the brain poses a challenge due to the restrictive nature of the blood-brain barrier (BBB). Here we report the development of mRNA-loaded lipid nanoparticles (LNPs) functionalized with BBB-interacting small molecules, thereby enhancing brain delivery and gene expression. Screening brain-targeted mRNA-LNPs in central nervous system (CNS) <i>in vitro</i> models and through intravenous administration in mice demonstrated that acetylcholine-conjugated LNPs achieved superior brain tropism and gene expression, outperforming LNP modifications with nicotine, glucose, memantine, cocaine, tryptophan, and other small molecules. An artificial intelligence (AI)-based model designed to predict the BBB permeability of small-molecule ligands showed strong alignment with our experimental results, providing <i>in vivo</i> validation of its predictive capacity. Cell-specific biodistribution analysis in Cre-reporter Ai9 mice showed that acetylcholine-functionalized LNPs preferentially transfected neurons and astrocytes following either intravenous or intracerebral administration. Mechanistic studies suggest that acetylcholine-LNP uptake is mediated by the functional engagement of acetylcholine receptors (AchRs) followed by endocytosis, which synergistically enhances intracellular mRNA delivery. Moreover, acetylcholine-LNPs successfully crossed a human BBB-on-a-chip model, enabling transgene expression in human iPSC-derived neurons. Their effective penetration and transfection in human brain organoids further support their potential activity in human-based systems. These findings establish a predictive and modular framework for engineering CNS-targeted LNPs, advancing precision gene delivery for brain disorders.

ACS NanoVol. 19(41)
Israel Institute for Biological Research (IL), Tel Aviv University (IL), Technion – Israel Institute of Technology (IL), Medical University of Graz (AT), University of Zurich (CH), Center for NanoScience (DE), Department of Biotechnology (IN), Institute for Stem Cell Biology and Regenerative Medicine (IN), Centre de Nanosciences et de Nanotechnologies (FR), Centre for Nano and Soft Matter Sciences (IN), Rappaport Family Institute for Research in the Medical Sciences (IL), Guangdong Technion-Israel Institute of Technology (CN), 3T Ophthalmics (United States) (US), Institute of Neurobiology (BG), National Assembly (KR), Institute of Pharmacology (CZ)
Openalex Percentile: Top 1%
ATP Synthase and ATPases Research
14.51
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