Expression of human F.IX as a function of time in plasma of Rag-1 mice following intramuscular injection of AAV vector at day 0.

(A) Systemic expression of human F.IX from CMV IE enhancer/promoter following intramuscular injection of AAV vectors into Rag-1 mice (n = 4 for each vector, 1 × 1011 vector genomes/mouse). Data points are mean expression levels at each time point, and vertical lines indicate standard deviation. AAV-CMV-hF.IX-2: open squares; AAV-CMV-hF.IX-3: open diamonds; AAV-CMV-hF.IX-3/bGH: open circles; AAV-CMV-hF.IX-8: open triangles. (B) Systemic expression of human F.IX from muscle-specific promoters (HSA or troponin) following intramuscular injection of AAV vector into Rag-1 mice (n = 4 for each vector, 1 × 1011 vector genomes/mouse). AAV-HSA-hF.IX-1: closed circle; AAV-HSA-hF.IX-3: closed square; AAV-cTnT-hF.IX: closed triangle. (C) Effect of HSA enhancer on systemic expression of human F.IX following intramuscular injection of AAV vector into Rag-1 mice (n = 4 for each vector, 1 × 1011 vector genomes/mouse). Note that vector AAV-HSA/CMV-hF.IX-2 (HSA/CMV-2, half-closed square) was constructed by the addition of a 1.2-kb 5′-flanking sequence of the HSA promoter 5′ to the CMV IE enhancer/promoter in vector AAV-CMV-hF.IX-2 (CMV-2, open square). The 3′-UT region of the human F.IX cDNA was deleted in the vector to accommodate the additional sequence. Animals injected with AAV-CMV-hF.IX-2 were identical to those shown in graph A. (D) Summary of systemic human F.IX expression in Rag-1 mice from AAV vectors. Columns represent average values of mean human F.IX expression levels for each vector for time points 6 to 12 weeks postinjection, ie, the plateau of expression (average of data points shown in graphs A-C). Vertical bars are standard deviation from the mean.