Primate-conserved carbonic anhydrase IV and murine-restricted LY6C1 enable blood-brain barrier crossing by engineered viral vectors

• DOI: 10.1126/sciadv.adg6618
• Publication Date: 2023-04-19T17:58:40Z
• Authors: Timothy F. Shay; Erin E. Sullivan; Xiaozhe Ding; Xinhong Chen; Sripriya Ravindra...; David Goertsen; David Brown; Anaya Crosby; Jost Vielmetter; Máté Borsos; Damien A. Wolfe; Annie W. Lam; Viviana Gradinaru
• Keywords: Transcytosis

The blood-brain barrier (BBB) presents a major challenge for delivering large molecules to study and treat the central nervous system. This is due in part scarcity of targets known mediate BBB crossing. To identify novel targets, we leverage panel adeno-associated viruses (AAVs) previously identified through mechanism-agnostic directed evolution improved transcytosis. Screening potential cognate receptors enhanced crossing, two targets: murine-restricted LY6C1 widely conserved carbonic anhydrase IV (CA-IV). We apply AlphaFold-based silico methods generate capsid-receptor binding models predict affinity AAVs these receptors. Demonstrating how tools can unlock target-focused engineering strategies, create an LY6C1-binding vector, AAV-PHP.eC, that, unlike our prior PHP.eB, also works Ly6a-deficient mouse strains such as BALB/cJ. Combined with structural insights from computational modeling, identification primate-conserved CA-IV enables design more specific potent human brain-penetrant chemicals biologicals, including gene delivery vectors.