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bioRxiv · 10.64898/2026.03.15.711945

LRP6-Guided Engineering of AAV9 Variants with Enhanced Blood-Brain Barrier Penetration and Reduced Liver Tropism in Non-Human Primates

Wang, Z.·Xu, X.·Sun, Z.·Li, H.·He, R.·Xu, Y.·Yu, M.·Wang, S.·Hu, C.·Liu, L.·Ren, L.·Zhang, L.·Xiao, T.·Luo, Y.·An, Z.
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Abstract

The blood-brain barrier (BBB) severely restricts the delivery of systemically administered adeno-associated virus (AAV) vectors for central nervous system (CNS) gene therapy. To overcome this limitation, we engineered a library of AAV9 capsid variants through rational design focused on the low-density lipoprotein receptor-related protein 6 (LRP6), a conserved mediator of transcytosis. A multi-tiered screening strategy, encompassing human BBB endothelial cells followed by neuronal and glial target cells in vitro, identified three lead variants (QL9-21, QL9-22, and QL9-25) with markedly enhanced transduction potential. In mice, these variants achieved a 5-28 fold increase in brain-wide gene delivery compared to AAV9, without elevating hepatic tropism. Crucially, evaluation in non-human primates (NHPs) revealed that the lead variant, QL9-21, mediated a striking 3-40 fold enhancement in viral genome delivery across all examined brain regions versus AAV9, while concurrently reducing liver accumulation by 2.6 fold. Our study establishes an LRP6-guided engineering platform that yields novel AAV9 vectors capable of efficient, species-conserved BBB penetration coupled with a favorable safety profile, representing a significant advance toward clinically translatable CNS gene therapies.

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Wang, Z., Xu, X., Sun, Z., Li, H., He, R., Xu, Y., Yu, M., Wang, S., Hu, C., Liu, L., Ren, L., Zhang, L., Xiao, T., Luo, Y., An, Z.. 2026-03-18. LRP6-Guided Engineering of AAV9 Variants with Enhanced Blood-Brain Barrier Penetration and Reduced Liver Tropism in Non-Human Primates. https://doi.org/10.64898/2026.03.15.711945

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