Rapamycin control of exocrine protein levels in saliva after adenoviral vector-mediated gene transfer.
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 microg/ml. Over a 16-day experimental period, three separate administrations of rapamycin (3 mg/kg) induced distinct elevations of salivary hGH (approximately 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.
Wang
J
J
Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, DHHS, Bethesda, MD 20892-1190, USA.
Voutetakis
A
A
Zheng
C
C
Baum
B J
BJ
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000900:Anti-Bacterial Agents / Q000627:therapeutic use*; D002460:Cell Line; D015870:Gene Expression / Q000187:drug effects; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage; D019382:Human Growth Hormone / Q000032:analysis* / Q000235:genetics; D006801:Humans; D019226:Oral Ulcer / Q000628:therapy*; D010862:Pilocarpine; D051381:Rats; D012463:Saliva / Q000737:chemistry*; D020123:Sirolimus / Q000627:therapeutic use*; D014161:Transduction, Genetic / Q000379:methods; D019076:Transgenes
D000900:Anti-Bacterial Agents; D010862:Pilocarpine; D019382:Human Growth Hormone; D020123:Sirolimus