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PreprintCited by 67Barrier Structure and Function StudiesVirus-based gene therapy researchRNA Interference and Gene Delivery

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Primate-conserved Carbonic Anhydrase IV and murine-restricted Ly6c1 are new targets for crossing the blood-brain barrier

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Shay, Timothy F., Sullivan, Erin E., Ding, Xiaozhe, Chen, Xinhong, Kumar, Sripriya Ravindra, Goertsen, David, Brown, David, Vielmetter, Jost, Borsos, Máté, Lam, Annie W., Gradinaru, Viviana·2023·Science Advances
DOI 10.1101/2023.01.12.523632PMC PMC10115422

Preprint — published as 10.1126/sciadv.adg6618

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Abstract

The blood-brain barrier (BBB) presents a major challenge to delivering large molecules to study and treat the central nervous system (CNS). This is due in part to the scarcity of effective targets for BBB crossing, the identification of which is the crucial first step of drug development. Here, we leveraged a panel of adeno-associated viruses (AAVs) previously identified through directed evolution for improved BBB transport to reverse engineer protein targets for enhanced BBB crossing. We identify both murine-restricted Ly6c1 and primate-conserved carbonic anhydrase IV (Car4; CA4) as novel receptors for crossing the BBB. We demonstrate how these receptors can unlock new experimental and computational target-focused engineering strategies by creating the enhanced Ly6c1-binding vector AAV-PHP.eC and by applying AlphaFold2-enabled in silico methods to rank capsids against identified receptors and generate capsid-receptor binding models. Here, with Car4, we add a completely new receptor to the very short list currently available for crossing the BBB in humans and, with Ly6c1, we validate a pipeline for receptor-targeted engineering. The identification of Car4/CA4 and structural insights from computational modeling provide new paths toward human brain-penetrant chemicals (drugs) and biologicals (including gene delivery).

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Metadata

Authors
Shay, T. F., Sullivan, E. E., Ding, X., Chen, X., Kumar, S. R., Goertsen, D., Brown, D., Vielmetter, J., Borsos, M., Lam, A. W., & Gradinaru, V.
Affiliations
California Institute of Technology (US)
Corresponding author
  • Shay, Timothy F.
  • Sullivan, Erin E.
  • Ding, Xiaozhe
  • Gradinaru, Viviana
ORCID
Venue
Science Advances
Publisher
openRxiv
Published
2023-01-13
Volume
9
Issue
16
Pages
eadg6618-eadg6618
ISSN
2375-2548 (ISSN-L)
DOI
10.1101/2023.01.12.523632
PMC
PMC10115422
OpenAlex
W4366383240
Cited by
67
References
1
Citations by year
2026: 24 · 2025: 25 · 2024: 13 · 2023: 4
FWCI
6.78
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Science Advances (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • California Institute of Technology
  • National Institutes of Health (DP1OD025535, 1uf1mh128336-01)
  • National Institute of Mental Health (U24MH131054, UF1 MH128336)
  • National Institute of Neurological Disorders and Stroke (DP1NS111369)
Topics
  • Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Keywords
blood-brain barrier penetration, adeno-associated virus vectors, carbonic anhydrase IV, directed evolution, brain drug delivery
MeSH terms
Dependovirus, Animals, Blood-Brain Barrier, Brain, Humans, Primates, Gene Transfer Techniques, Carbonic Anhydrase IV, Mice

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