In vivo selection in non-human primates identifies AAV capsids for on-target CSF delivery to spinal cord.

Publisher: Cell Press Country of Publication: United States NLM ID: 100890581 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1525-0024 (Electronic) Linking ISSN: 15250016 NLM ISO Abbreviation: Mol Ther Subsets: MEDLINE

Publication: 2017- : Cambridge, MA : Cell Press
Original Publication: San Diego, CA : Academic Press, 2000-

Dependovirus*/genetics ; Spinal Cord*/metabolism ; Genetic Vectors*/administration & dosage ; Genetic Vectors*/genetics ; Genetic Therapy*/methods; Animals ; Humans ; Transgenes ; Gene Transfer Techniques ; Capsid/metabolism ; Tissue Distribution ; Injections, Spinal ; Transduction, Genetic ; Macaca mulatta ; Capsid Proteins/genetics ; Capsid Proteins/metabolism

Systemic administration of adeno-associated virus (AAV) vectors for spinal cord gene therapy has challenges including toxicity at high doses and pre-existing immunity that reduces efficacy. Intrathecal (IT) delivery of AAV vectors into cerebral spinal fluid can avoid many issues, although distribution of the vector throughout the spinal cord is limited, and vector entry to the periphery sometimes initiates hepatotoxicity. Here we performed biopanning in non-human primates (NHPs) with an IT injected AAV9 peptide display library. We identified top candidates by sequencing inserts of AAV DNA isolated from whole tissue, nuclei, or nuclei from transgene-expressing cells. These barcoded candidates were pooled with AAV9 and compared for biodistribution and transgene expression in spinal cord and liver of IT injected NHPs. Most candidates displayed increased retention in spinal cord compared with AAV9. Greater spread from the lumbar to the thoracic and cervical regions was observed for several capsids. Furthermore, several capsids displayed decreased biodistribution to the liver compared with AAV9, providing a high on-target/low off-target biodistribution. Finally, we tested top candidates in human spinal cord organoids and found them to outperform AAV9 in efficiency of transgene expression in neurons and astrocytes. These capsids have potential to serve as leading-edge delivery vehicles for spinal cord-directed gene therapies.
Competing Interests: Declaration of interests C.A.M. has financial interests in Chameleon Biosciences, Skylark Bio, and Sphere Gene Therapeutics, companies developing adeno-associated virus (AAV) vector technologies for gene therapy applications. C.A.M. performs paid consulting work for all three companies. C.A.M. received sponsored research funding from SwanBio Therapeutics for the research described here. C.A.M. received royalty payments from licensing agreements between SwanBio Therapeutics and the Massachusetts General Hospital. C.A.M.’s interests were reviewed and are managed by Massachusetts General Hospital and Mass General Brigham in accordance with their conflict-of-interest policies. C.A.M. and K.S.H. have a filed patent application surrounding the iTransduce library. K.S.H. performed paid consulting work for SwanBio Therapeutics.
(Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Update of: bioRxiv. 2023 Sep 13:2023.09.13.557506. doi: 10.1101/2023.09.13.557506. (PMID: 37745398)

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Keywords: AAV; AAV capsid libraries; adeno-associated virus; engineered AAV capsids; gene therapy; intrathecal injection; lumbar injection; spinal cord; spinal cord organoids; viral vector

0 (Capsid Proteins)

Date Created: 20240607 Date Completed: 20240808 Latest Revision: 20240921

20240921

PMC11405149

10.1016/j.ymthe.2024.05.040

38845196