Adeno-associated virus serotype 9 vectors transduce murine alveolar and nasal epithelia and can be readministered

doi:10.1073/pnas.0601433103

Adeno-associated virus serotype 9 vectors transduce murine alveolar and nasal epithelia and can be readministered

Maria P. Limberis and
James M. Wilson*

Gene Therapy Program, Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104

Edited by Inder M. Verma, The Salk Institute for Biological Studies, La Jolla, CA, and approved July 10, 2006 (received for review March 22, 2006)

Abstract

Airway-directed gene transfer has emerged as a promising approach for the treatment of the two genetic diseases of the lung, namely cystic fibrosis and α-1-antitrypsin deficiency. Herein we describe the transduction efficiency of a novel adeno-associated virus (AAV) vector, AAV2/9, across murine nasal and lung airway epithelia. At the peak of gene expression AAV2/9-mediated human α-1-antitrypsin gene expression in serum was ≈60-fold better than that of AAV2/5. We found that AAV2/9-mediated nLacZ gene transfer in nasal and lung airways was relatively stable for 9 months, suggesting that a progenitor airway cell population was transduced. Most interestingly, we show that AAV2/9 can be readministered in the presence of high levels of serum-circulating neutralizing antibodies as early as 1 month after initial exposure, with minimal effect on overall reporter gene expression, rendering it a promising gene transfer vector candidate for use in humans.

Footnotes

*To whom correspondence should be addressed at:
Translational Research Laboratory, 125 South Thirty-First Street, Suite 2000, Philadelphia, PA 19104-3403.
E-mail: wilsonjm{at}mail.med.upenn.edu
Author contributions: M.P.L. and J.M.W. designed research; M.P.L. performed research; M.P.L. and J.M.W. analyzed data; and M.P.L. and J.M.W. wrote the paper.
Conflict of interest statement: J.M.W. is an inventor on patents describing these vectors, which have been licensed.
This paper was submitted directly (Track II) to the PNAS office.
Abbreviations::

AAV,

adeno-associated virus;

AAT,

α-1-antitrypsin;

hAAT,

human AAT;

CF,

cystic fibrosis;

NAB,

neutralizing antibody;

i.t.,

intratracheal(ly);

i.n.,

intranasal(ly);

BALF,

bronchoalveolar lavage fluid;

SNK,

Student–Newman–Keuls;

GC,

genome copies.