Sorting behavior of a transgenic erythropoietin-growth hormone fusion protein in murine salivary glands.
Salivary glands are useful gene transfer target sites for the production of therapeutic proteins, and can secrete proteins into both saliva and the bloodstream. The mechanisms involved in this differential protein sorting are not well understood, although it is believed, at least in part, to be based on the amino acid sequence of the encoded protein. We hypothesized that a transgenic protein, human erythropoietin (hEpo), normally sorted from murine salivary glands into the bloodstream, could be redirected into saliva by fusing it with human growth hormone (hGH). After transfection, the hEpo-hGH fusion protein was expressed and glycosylated in both HEK 293 and A5 cells. When packaged in an adenovirus serotype 5 vector and delivered to murine submandibular cells in vivo via retroductal cannulation, the hEpo-hGH fusion protein was also expressed, albeit at approximately 26% of the levels of hEpo expression. Importantly, in multiple experiments with different cohorts of mice, the hEpo-hGH fusion protein was sorted more frequently into saliva, versus the bloodstream, than was the hEpo protein (p < 0.001). These studies show it is possible to redirect the secretion of a transgenic constitutive pathway protein from salivary gland cells after gene transfer in vivo, a finding that may facilitate developing novel treatments for certain upper gastrointestinal tract disorders.
Samuni
Yuval
Y
Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Department of Health and Human Services, Bethesda, MD 20892, USA.
Cawley
Niamh X
NX
Zheng
Changyu
C
Cotrim
Ana P
AP
Loh
Y Peng
YP
Baum
Bruce J
BJ
D000818:Animals; D002460:Cell Line; D004921:Erythropoietin / Q000235:genetics / Q000378:metabolism*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D019382:Human Growth Hormone / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008297:Male; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012469:Salivary Glands / Q000378:metabolism*; D014161:Transduction, Genetic; D019076:Transgenes
D011993:Recombinant Fusion Proteins; D004921:Erythropoietin; D019382:Human Growth Hormone