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Activators of viral gene expression in polarized epithelial monolayers identified by rapid-throughput drug screening

Gene Therapy volume 13, pages 781–788 (2006)Cite this article

Abstract

Epithelial polarity and tight junction formation limit the ability of adenovirus, retrovirus and adeno-associated virus (AAV) to deliver and express virally encoded genes. Using an extended half-life luciferase assay and high-throughput luminometry, we screened 23 000 compounds and natural product extracts as potentiators to overcome this barrier. Seven strong activators were discovered (up to several hundred fold above control) and two of these exhibited spectrum of activity in multiple cell types (HeLa (human cervical carcinoma), cystic fibrosis bronchial epithelial (human bronchial), HT29 (human colonic carcinoma), Calu3 (airway serous glandular)). Enhanced transduction by unrelated gene transfer vectors (adenovirus, lentivirus, AAV, liposomal) was also observed. These results establish a strategy for identifying compounds that improve viral gene transfer to resistant cell types, and provide new tools for examining epithelial defense against viral infection. The compounds should have broad usefulness in experimental therapies for cancer and genetic diseases.

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Acknowledgements

We are grateful to Mrs Jane Schell for typing the manuscript. Collections of Antarctic marine invertebrates and preparation of their extracts were supported by National Science Foundation Grants OPP-9530735 (McClintock), OPP-9626610 (Baker), OPP-9814538 (McClintock and Amsler) and OPP-9901076 (Baker). This work was supported by NIH (U19 CA67763, P30 DK54781) and the Cystic Fibrosis Foundation (R464-CR02). We thank Drs Jill Johnson and Robert Schultz from the NCI Developmental Therapeutics Program for providing us with the Diversity Set.

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Author notes

  1. E J Sorscher and J Harris: These two authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA

    E J Sorscher

  2. Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL, USA

    E J Sorscher, J Harris, M Alexander, A Rottgers, K Hardy & J S Hong

  3. Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA

    S Ponnazhagan

  4. Department of Cell Biology and Anatomy, University of Alabama at Birmingham, Birmingham, AL, USA

    J F Collawn & J S Hong

  5. Department of Biology, University of Alabama at Birmingham, Birmingham, AL, USA

    J McClintock & C D Amsler

  6. Department of Pharmaceutical Sciences, Samford University, Birmingham, AL, USA

    A Webster

  7. Southern Research Institute, Birmingham, AL, USA

    J Maddry

  8. Department of Chemistry, University of South Florida, Tampa, FL, USA

    B J Baker

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Correspondence to J S Hong.

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Sorscher, E., Harris, J., Alexander, M. et al. Activators of viral gene expression in polarized epithelial monolayers identified by rapid-throughput drug screening. Gene Ther 13, 781–788 (2006). https://doi.org/10.1038/sj.gt.3302676

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