A3 - AAV Vectors - Capsid Engineering
991: Human CA4-Targeting AAV Capsids Efficiently Transduce the CNS in Humanized Mice
Type: Poster Session
Poster Board Number: 991
Presentation Details
Session Title: Thursday Posters: AAV Vectors - Capsid Engineering
The development of adeno-associated viruses (AAVs) that efficiently target the human central nervous system (CNS) promises to boost the impact of gene therapies for neurological disorders. We engineered a suite of peptide-modified AAV9 capsids to cross the blood-brain barrier (BBB) by directly binding to the human GPI-linked enzyme Carbonic anhydrase IV (hCA4). Using our recently described receptor-targeting approach, we identified three capsids with distinct sequences, BI-hCA4-1, BI-hCA4-2, and BI-hCA4-3, that more efficiently transduce hCA4-expressing cells compared to wild type AAV9. When systemically injected into mice that express hCA4 on the brain microvasculature, the BI-hCA4 capsids transduced cells throughout the CNS more efficiently than AAV9 (Fig. 1). This enhanced tropism was not observed in wild type mice. We are now using site-saturation mutagenesis and a generative machine learning model paired with an AAV production fitness model to further improve the ability of the capsids to bind hCA4 and cross the BBB
in vivo. In summary, we demonstrate that human CA4 is a promising new target for engineering CNS-directed gene delivery vectors.
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Figure 1. Human CA4-targeted capsids achieve efficient gene delivery to the CNS. The hCA4 gene was first delivered to the CNS endothelium by intravenous administration of AAV-BI30:CAG-hCA4-miR122-WPRE. Next, AAV9 or BI-hCA4:CAG-mScarlet-P2A-Luciferase-SV40pA were administered intravenously. Representative images show AAV transduction in sagittal brain sections of mice three weeks after the delivery of the hCA4-binding AAV.
Nuria Roxana Botticello-Romero, Jason Wu
, Qin Huang
, Thomas Beddow
, Ken Y. Chan
, Pamela Brauer
, Fatma Elzahraa Eid
, Alina Chan
, Benjamin E. Deverman
Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA"