Evaluation of a rapamycin-regulated serotype 2 adeno-associated viral vector in macaque parotid glands.
OBJECTIVE
Salivary glands are useful target organs for local and systemic gene therapeutics. For such applications, the regulation of transgene expression is important. Previous studies by us in murine submandibular glands showed that a rapamycin transcriptional regulation system in a single serotype 2, adeno-associated viral (AAV2) vector was effective for this purpose. This study evaluated if such a vector was similarly useful in rhesus macaque parotid glands.


METHODS
A recombinant AAV2 vector (AAV-TF-RhEpo-2.3w), encoding rhesus erythropoietin (RhEpo) and a rapamycin-inducible promoter, was constructed. The vector was administered to macaques at either of two doses [1.5 x 10(11) (low dose) or 1.5 x 10(12) (high dose) vector genomes] via cannulation of Stensen's duct. Animals were followed up for 12-14 weeks and treated at intervals with rapamycin (0.1 or 0.5 mg kg(-1)) to induce gene expression. Serum chemistry, hematology, and RhEpo levels were measured at interval.


RESULTS
AAV-TF-RhEpo-2.3w administration led to low levels of rapamycin-inducible RhEpo expression in the serum of most macaques. In five animals, no significant changes were seen in serum chemistry and hematology values over the study. One macaque, however, developed pneumonia, became anemic and subsequently required euthanasia. After the onset of anemia, a single administration of rapamycin led to significant RhEpo production in this animal.


CONCLUSIONS
Administration of AAV-TF-RhEpo-2.3w to macaque parotid glands was generally safe, but led only to low levels of serum RhEpo in healthy animals following rapamycin treatment.
Zheng
C
C
Molecular Physiology and Therapeutics Branch, National Institute of Dental and Craniofacial Research, Bethesda, MD 20892-1190, USA.
Voutetakis
A
A
Metzger
M
M
Afione
S
S
Cotrim
A P
AP
Eckhaus
M A
MA
Rivera
V M
VM
Clackson
T
T
Chiorini
J A
JA
Donahue
R E
RE
Dunbar
C E
CE
Baum
B J
BJ
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D004305:Dose-Response Relationship, Drug; D004921:Erythropoietin / Q000097:blood / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D005822:Genetic Vectors / Q000008:administration & dosage*; D008253:Macaca mulatta; D008297:Male; D010306:Parotid Gland / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins; D020123:Sirolimus / Q000494:pharmacology*; D014161:Transduction, Genetic; D019076:Transgenes
D011994:Recombinant Proteins; D004921:Erythropoietin; D020123:Sirolimus