Clinical improvement after treatment with VEGF(165) in patients with severe chronic lower limb ischaemia

Abstract

The present study focuses on the application of a therapeutic strategy in patients with chronic severe lower limb ischaemia using a plasmid vector encoding the vascular endothelial growth factor (phVEGF(165)). It has been shown that VEGF promotes neo-vascularization and blood vessel network formation and thus might have the ability to improve blood-flow at the level of the affected limbs. However, little information is available regarding the necessary level of expression of VEGF and its possible related adverse effects. We have subcloned VEGF ( 165 )isoform into pCMV-Script expression vector (Stratagene) under the control of the CMV promoter. Three patients with chronic ischaemia of the lower limb, considered as not suitable for surgical re-vascularization, received intramuscular injection with 0.5 ml saline solution containing 10(11) copies of VEGF ( 165 ) plasmid. The clinical evolution has been monitored by angiography and estimated by walking time on the rolling carpet (Gardner protocol). Two months after therapy, all three patients showed complete relief of rest pain, improvement of ischaemic ulcer lesions and increased walking distance on the rolling carpet most probably due to appearance of newly formed collateral vessels.

Fig. 1

The injection sites

Fig. 2

Changes in C-Reactive Protein value assessed before (baseline) and after (6 h and 2 months) gene therapy

Fig. 3

Changes in erythrocyte sedimentation rate value (mm/h) assessed before (baseline) and after (6 h and 2 months) gene therapy

Fig. 4

Patient with chronic lower limb ischaemia. Pictures were taken on day 0 (left) and 2 months after VEGF gene therapy (right)

Fig. 5

Chart representing the walking test results for the 3 patients before gene therapy (the first symbol on each coloured line), after 1 month (the second symbol on each coloured line) and 2 months (the third symbol on each coloured line), respectively

Fig. 6

Angiographic assessment of collateral vessel formation in 1 patient. Compared to the angiography performed before VEGF gene therapy, new developed collateral vessels were visibile at the calf level two months after plasmid injection (see the encircled area)