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Rapamycin control of exocrine protein levels in saliva after adenoviral vector-mediated gene transfer

Gene Therapy volume 11, pages 729–733 (2004)Cite this article

Abstract

Transgene-encoded therapeutic secretory proteins can be efficiently secreted from salivary glands into saliva or the bloodstream after adenoviral (Ad)-mediated gene transfer. Since transgene expression from conventional vectors is typically unregulated, we evaluated the rapamycin-based dimerizer regulation system for control of transgene expression in, and consequent exocrine protein secreted from, rat salivary glands. We used human growth hormone (hGH) as a surrogate exocrine secretory protein. Two Ad vectors, Ad C4ZF3, encoding activation and DNA binding domain fusion polypeptides, and Ad Z12-I-GH-2, encoding hGH, were constructed and shown useful in vitro. Thereafter, both vectors were delivered into submandibular glands by retroductal infusion. After 24 h, rapamycin (0, 1, 3 or 10 mg/kg) was administered, and 20 h later hGH levels in saliva were determined. Salivary hGH levels were rapamycin concentration dependent. At a rapamycin dose of 10 mg/kg, total salivary hGH was 693±197 ng and the hGH concentration in saliva was 4.6±1.3 μg/ml. Over a 16-day experimental period, three separate administrations of rapamycin (3 mg/kg) induced distinct elevations of salivary hGH (∼100–200 ng total hGH) that were entirely rapamycin dependent. This study demonstrates for the first time pharmacological control of transgenic exocrine protein production and presence in saliva after salivary gland gene transfer, and the potential for its application to the management of oral, oropharyngeal and upper gastrointestinal tract disorders.

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Acknowledgements

We thank Dr Victor Rivera and ARIAD Pharmaceuticals (www.ariad.com/regulationkits) for providing the plasmids, rapamycin and AP21967 used in these experiments. We also thank Dr Rivera, and Drs J Chiorini, A Cotrim, M Schmidt and R Wellner for their comments on an earlier draft of this manuscript.

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Authors and Affiliations

  1. Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, DHHS, Bethesda, MD, USA

    J Wang, A Voutetakis, C Zheng & B J Baum

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  1. J Wang
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  2. A Voutetakis
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  4. B J Baum
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Wang, J., Voutetakis, A., Zheng, C. et al. Rapamycin control of exocrine protein levels in saliva after adenoviral vector-mediated gene transfer. Gene Ther 11, 729–733 (2004). https://doi.org/10.1038/sj.gt.3302225

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