A novel adenoviral gutless vector encoding sphingosine kinase promotes arteriogenesis and improves perfusion in a rabbit hindlimb ischemia model

Objectives: We previously demonstrated that sphingosine kinase (SPK) increases the level of extracellular sphingosine-1-phosphate and promotes neovascularization in a mouse matrigel model. In this study, we tested the hypothesis that SPK gene transfer using a novel adenoviral 'gutless' vector (AGV) can enhance arteriogenesis in a rabbit hindlimb ischemia model. Methods: Thirty-five male New Zealand white rabbits were randomized to the AGV-SPK group (n = 13), AGV-null group (n = 13), and control group (n = 9). On day 10, after the induction of unilateral hindlimb ischemia, gene vectors or buffer were introduced and the effect examined on day 30, using calf blood pressure, quantitative angiographic analysis, and histology. Results: Calf systolic blood pressure ratios of the ischemic limb to the normal limb on day 30 were 0.77 ± 0.13 in control groups, including the AGV-null group, and 0.91 ± 0.14 in the AGV-SPK group (P < 0.05). Angiographic vessel counts were significantly increased (8.0 ± 2.1 at baseline and 11.8 ± 3.2 on day 30, P < 0.001) in the AGV-SPK group. Histologic analysis showed that microscopic total vessel counts on day 30 were 3.5 ± 1.8/field in the control and AGV-null group and 5.4 ± 1.0/field in the AGV-SPK group. Arterioles (AGV-SPK; 3.0 ± 0.8 versus control and AGV-null; 2.1 ± 1.1, P < 0.05) were significantly increased in the AGV-SPK group. Conclusions: This study shows that SPK promotes arteriogenesis, as evidenced by the maximal improvement in the blood pressure restoration and collateral vessel counts. SPK may be an important angiogenic target to improve perfusion in ischemic tissues.