Control of erythropoietin delivery by doxycycline in mice after intramuscular injection of adeno-associated vector.
We reported previously that controlled expression of a foreign gene in response to tetracycline derivative can be accomplished in mice by the autologous transplantation of retrovirus-modified muscle cells. Although regulated systemic delivery of therapeutic proteins from engineered tissues has potential clinical application, the transplantation of muscle cells is not currently feasible in humans. Several studies have shown that a single injection of adeno-associated virus (AAV) vectors into mouse muscle results in long-term expression of reporter genes as well as sustained delivery of proteins into the serum. Because this method is potentially applicable clinically, we constructed an AAV vector in which the expression of the mouse erythropoietin (Epo) cDNA is modulated in response to doxycycline. The vector was injected intramuscularly in normal mice. We observed that hematocrit and serum Epo concentrations could be modulated over a 29-week period in response to the presence or absence of doxycycline in the drinking water of these animals. Thus, a regulated gene expression cassette can be incorporated into a single AAV vector, such that intramuscular injection of the vector allows sustained and regulated expression of a desired gene.
Bohl
D
D
Laboratoire Rétrovirus et Transfert Génétique, CNRS URA 1157, Institut Pasteur, Paris, France.
Salvetti
A
A
Moullier
P
P
Heard
J M
JM
D000818:Animals; D015152:Blotting, Northern; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000008:administration & dosage*; D000229:Dependovirus / Q000235:genetics*; D004318:Doxycycline / Q000008:administration & dosage / Q000494:pharmacology*; D004921:Erythropoietin / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D007273:Injections, Intramuscular; D051379:Mice; D009132:Muscles / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis
D018076:DNA, Complementary; D012333:RNA, Messenger; D004921:Erythropoietin; D004318:Doxycycline