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Polarized AAVR expression determines infectivity by AAV gene therapy vectors

Gene Therapy (2019) | Download Citation


Adeno-associated virus (AAV) has been investigated to transfer the cystic fibrosis transmembrane conductance regulator (CFTR) to airways. Inhaled AAV2-CFTR in people with cystic fibrosis (CF) is safe, but inefficient. In vitro, AAV2 transduction of human airway epithelia on the apical (luminal) side is inefficient, but efficient basolaterally. We previously selected AAV2.5T, a novel capsid that apically transduces CF human airway epithelia and efficiently restores CFTR function. We hypothesize the AAV receptor (AAVR) is basolaterally localized, and that AAV2.5T utilizes an alternative apical receptor. We found AAVR in human airway epithelia by western blot and RNA-Seq analyses. Using immunocytochemistry we did not find endogenous AAVR at membranes but overexpression localized AAVR to the basolateral membrane, where it preferentially increased transduction. Anti-AAVR antibodies blocked transduction by AAV2 from the basolateral side but not AAV2.5T from the apical side, suggesting a unique apical receptor. Finally, we found infection by AAV2 but not AAV2.5T was blocked by CRISPR knockout of AAVR in cell lines. Our data suggest the absence of apical AAVR is rate limiting for AAV2, and efficient transduction by AAV2.5T is accomplished using an AAVR independent pathway. Our findings inform the development of gene therapy for CF, and AAV vectors in general.

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This work was supported by grants from the NIH (H678200-G) and the University of Iowa Center for Gene Therapy (DK054759).

Author information


  1. Department of Internal Medicine, Roy J. and Lucille A. Carver College of Medicine, Pappajohn Biomedical Institute, The University of Iowa, Iowa City, IA, USA

    • Bradley A. Hamilton
    • , Xiaopeng Li
    • , Alejandro A. Pezzulo
    • , Mahmoud H. Abou Alaiwa
    •  & Joseph Zabner
  2. Molecular Medicine Program, The University of Iowa, Iowa City, IA, USA

    • Bradley A. Hamilton
    •  & Joseph Zabner
  3. Department of Biomedical Engineering, The University of Iowa, Iowa City, IA, USA

    • Mahmoud H. Abou Alaiwa


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Conflict of interest

Dr. Zabner is a founder and holds equity in TaleeBio. The other authors declare that they have no conflict of interest.

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Correspondence to Joseph Zabner.

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