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J. Biol. Chem., Vol. 281, Issue 40, 29684-29692, October 6, 2006

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Unique Biologic Properties of Recombinant AAV1 Transduction in Polarized Human Airway Epithelia^*

Ziying Yan, Diana C. M. Lei-Butters, Xiaoming Liu, Yulong Zhang, Liang Zhang, Meihui Luo, Roman Zak, and John F. Engelhardt^¶1

From the Department of Anatomy and Cell Biology, ^¶Department of Internal Medicine, and Center for Gene Therapy of Cystic Fibrosis and Other Genetic Diseases, University of Iowa College of Medicine, Iowa City, Iowa 52242

The choice of adeno-associated virus serotypes for clinical applications is influenced by the animal model and model system used to evaluate various serotypes. In the present study, we sought to compare the biologic properties of rAAV2/1, rAAV2/2, and rAAV2/5 transduction in polarized human airway epithelia using viruses purified by a newly developed common column chromatography method. Results demonstrated that apical transduction of human airway epithelia with rAAV2/1 was 100-fold more efficient than rAAV2/2 and rAAV2/5. This transduction profile in human airway epithelia (rAAV2/1 >> rAAV2/2 = rAAV2/5) was significantly different from that seen following nasal administration of these vectors to mouse lung (rAAV2/5 > rAAV2/1 >> rAAV2/2), emphasizing differences in transduction of these serotypes between these two species. In stark contrast to rAAV2/2 and rAAV2/5, rAAV2/1 transduced both the apical and basolateral membrane of human airway epithelia with similar efficiency. However, the overall level of transduction across serotypes did not correlate with vector internalization. We hypothesized that differences in post-entry processing of these serotypes might influence the efficiency of apical transduction. To this end, we tested the effectiveness of proteasome inhibitors to augment nuclear translocation and gene expression from the three serotypes. Augmentation of rAAV2/1 apical transduction of human polarized airway epithelia was 10-fold lower than that for rAAV2/2 and rAAV2/5. Cellular fractionation studies demonstrated that proteasome inhibitors more significantly enhanced rAAV2/2 and rAAV2/5 translocation to the nucleus than rAAV2/1. These results demonstrate that AAV1 transduction biology in human airway epithelia differs from that of AAV2 and AAV5 by virtue of altered ubiquitin/proteasome sensitivities that influence nuclear translocation.

Received for publication, April 28, 2006 , and in revised form, July 10, 2006.

^* This work was supported by National Institutes of Health Grant RO1 HL58340 (to J. F. E.) Center for Gene Therapy Grant DK54759, and by the Targeted Genetics Corp. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed: Rm. 1-111 BSB, Dept. of Anatomy and Cell Biology, College of Medicine, University of Iowa, 51 Newton Rd., IA City, IA 52242. Tel.: 319-335-7744; Fax: 319-335-6581; E-mail: john-engelhardt{at}uiowa.edu.

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