Gene Therapy in Rats with a Lentiviral Vector Containing the Human Coagulation Factor IX Gene / 대한수혈학회지

ABSTRACT

BACKGROUND: Hemophilia B is an inheritable X-linked bleeding disorder that occurs as a consequence of genetic alterations within the factor IX (IX) gene. In the present study, pseudotyped HIV-I-derived lentiviral vectors expressing human IX (lentivirus-IX) were assessed for the ability to produce an active human IX in the animals transduced with lentivirus-IX. METHODS: The IX concentrations and activated partial thromboplastin times (aPTT) were measured from the supernatants of HeLa cells that were transduced with lentivirus-IX. In an animal study, we injected 1microgram of lentivirus-IX into the hind limbs of Sparague-Dawley (SD) rats. The IX concentrations were measured from the plasma of the vehicle injected rats and the plasma of the lentivirus-IX injected rats for 8 weeks. RESULTS: The in vitro expression of human IX was detected in a dose-dependent manner following the transduction of lentivirus-IX into the HeLa cells (control: 10+/-3 vs. 100 ng of lentivirus-IX: 1486+/-50 ng/mL, P<0.05). The aPTT also showed the tendency of dose-dependent decrease (control: 83.9+/-0.5 vs. 50 ng of lentivirus-IX: 80.1+/- 0.8 sec), but this was not statistically significant. In the animal experiment, the plasma IX concentration from the lentivirus-IX transduced rats (n=3) was significantly increased compared to the vehicle-injected rats (n=4) (5.9+/-3.9 vs. 46.4+/-20.6 ng/mL) at post-injection 1 week. CONCLUSION: This study demonstrated that in vivo delivery of lentiviral vectors expressing human IX to the muscle cells has the potential to be a therapeutic modality for hemophilia B.