Prophylactic gene therapy with human tissue kallikrein ameliorates limb ischemia recovery in type 1 diabetic mice.
Diabetes macro- and microvascular disease causes tissue hypoperfusion. This deficit, together with a failure to mount an adequate angiogenic response, might explain why vascular occlusion evolves more severely among diabetic patients. The present study investigated whether prophylactic gene therapy with human tissue kallikrein (hTK) may protect diabetic limbs from the consequences of supervening ischemia. Vehicle (saline) or an adenovirus carrying the gene for either hTK (Ad.hTK) or luciferase (Ad.Luc) was injected into left adductor muscles of streptozotocin-induced type 1 diabetic mice 2 weeks before operative occlusion of the ipsilateral femoral artery. Saline-injected nondiabetic mice served as controls. Hindlimb blood flow recovery was analyzed sequentially over the 2 weeks after ischemia induction. At necroscopy, microvessel density and endothelial cell proliferation and apoptosis were quantified in skeletal muscles. We found that limb perfusion recovery of saline-injected type 1 diabetic mice is delayed because of insufficient reparative neovascularization and excessive activation of endothelial cell apoptosis. By contrast, prophylactic Ad.hTK renewed the ability to mount an appropriate neovascularization response to ischemia, suppressed apoptosis, and upregulated endothelial nitric oxide synthase expression. Ultimately, correction of diabetic endotheliopathy by Ad.hTK allowed proper perfusion recovery as seen in nondiabetic mice. These discoveries disclose new therapeutic options for the treatment of diabetic complications.
Emanueli
Costanza
C
Experimental Medicine and Gene Therapy Section, Istituto Nazionale Biostrutture e Biosistemi, Alghero and Osilo, Italy. emanueli@yahoo.com
Graiani
Galliam
G
Salis
Maria B
MB
Gadau
Sergio
S
Desortes
Elisa
E
Madeddu
Paolo
P
D000818:Animals; D001160:Arterioles / Q000187:drug effects / Q000254:growth & development; D003921:Diabetes Mellitus, Experimental / Q000628:therapy; D003922:Diabetes Mellitus, Type 1 / Q000628:therapy*; D003925:Diabetic Angiopathies / Q000628:therapy*; D005121:Extremities / Q000098:blood supply; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D007511:Ischemia / Q000209:etiology / Q000628:therapy; D008156:Luciferases / Q000235:genetics; D008297:Male; D051379:Mice; D019001:Nitric Oxide Synthase / Q000235:genetics; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; D012016:Reference Values; D013997:Time Factors; D020840:Tissue Kallikreins / Q000235:genetics*
D008156:Luciferases; C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse; D020840:Tissue Kallikreins