Ultrasound molecular imaging of angiogenesis induced by mutant forms of hypoxia-inducible factor-1α

AimsTargeted point mutants of hypoxia-inducible factor-1α (HIF-1α) are potential optimal agents for angiogenesis therapy. Data are limited regarding the angiogenic response of HIF-1α mutants. We aimed to compare the angiogenic effect of wild-type and mutant HIF-1α by contrast ultrasound molecular imaging (UMI) of αv-integrin expression.Methods and resultsThe wild-type gene of human HIF-1α, a gene with double mutations (HIF-1α564/803), a gene with triple mutations (HIF-1α564/803/402), or the LacZ gene (control) was transfected into the ischaemic hind limbs of C57BL/6 mice using an adenovirus vector. The video intensity of microbubbles targeted to αv-integrins in the ischaemic limbs increased along with the number of point mutations of HIF-1α. Immunohistochemical expression of endothelial αv-integrins was higher in the mutant HIF-1α564/803/402 group than the other groups as was the density of both capillaries and arterioles in ischaemic muscle. Expression of both the mRNA and protein for HIF-1α and VEGF was significantly higher in the mutant HIF-1α564/803/402 group than in the other groups. The half-life of HIF-1α and VEGF mRNA was longer in HIF-1α mutant-transfected cells than in wild-type HIF-1α or LacZ-transfected cells.ConclusionHIF-1α mutants were more effective for enhancing angiogenesis in ischaemic muscle tissue than wild-type HIF-1α, and the response could be qualitatively evaluated by UMI of αv-integrins expression.