Abstract 15180: Adeno-HSPA12B Delivery Improves Neovascularization in a Murine Model of Hind Limb Ischemia
Objective: Molecular chaperons/heat shock proteins play a critical role in angiogenesis. HSPA12B is a member of the HSP70 subfamily 12 and is predominantly expressed in endothelial cells and required for angiogenesis. In the present study we aim to investigate its role in a murine model of hind limb ischemia.
Methods: Adult 8-12 week old C57Bl/6J mice were divided into two groups: (1) control (Ad.LacZ) and (2) Adeno-HSPA12B gene treatment group (Ad.HSPA12B). Both groups underwent right femoral artery ligation to create hind limb ischemia. Immediately after surgery, mice in treatment group received Ad.HSPA12B in a concentration of 1x109 PFU in both semimembranosus and gastrocnemius muscles of the right leg whereas the left leg was used as an internal control. The mice in control group received similar concentration of Ad.LacZ. The two groups underwent serial Laser Doppler imaging both pre-operatively and post-operatively for 28 days to assess blood perfusion in the hind-limbs. Immunohistochemistry and Immunofluorescence were performed on post-operative day 3 and 28.
Results: Ad.HSPA12B treated group showed a significantly increased perfusion ratio on postoperative day 21 [0.65±0.04 (n=12) vs. 0.50±0.04 (n=16); p < 0.05] and day 28 [0.71±0.06 (n=12) vs. 0.55±0.03 (n=16);p < 0.05] as compared to mice in the Ad.LacZ group. Immunohistochemical analysis documented increased capillary density [1413±70.25 (n=5) vs. 586±26.39 (n=6);p < 0.05], capillary myocyte ratio [1.81±0.16 (n=5) vs. 1.29±0.10 (n=6); p < 0.05] along with reduced fibrosis in Ad.HSPA12B group as compared to Ad.LacZ at post-operative day 28. Ad.HSPA12B treatment group showed increased [1.39 fold] VEGF expression by immunofluorescence analysis compared to Ad.LacZ group, three days after femoral artery ligation.
Conclusions: Taken together, our study demonstrates that targeted gene therapy with Ad.HSPA12B in the murine ischemic muscle enhances blood perfusion, reduces fibrosis and improves neovascularization via VEGF expression. We anticipate that this molecule can be a future potential target for clinical trials and subsequently drug therapy in peripheral vascular disease management.
Author Disclosures: M.T. Rishi: None. V. Selvaraju: None. J. Palestey: None. N. Maulik: None.
- © 2016 by American Heart Association, Inc.