Neovascularization of ischemic tissues by gene delivery of the extracellular matrix protein Del-1
Jingping Zhong, … , Nancy Boudreau, Judith A. Varner
Jingping Zhong, … , Nancy Boudreau, Judith A. Varner
Published July 1, 2003
Citation Information: J Clin Invest. 2003;112(1):30-41. https://doi.org/10.1172/JCI17034.
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Article Cardiology

Neovascularization of ischemic tissues by gene delivery of the extracellular matrix protein Del-1

Abstract

The ECM protein Del-1 is one of several novel ECM proteins that accumulate around angiogenic blood vessels in embryonic and tumor tissue and promote angiogenesis in the absence of exogenous growth factors. Del-1 expressed in mouse or rabbit ischemic hind-limb muscle by gene transfer rapidly promotes new blood vessel formation and restores muscle function. This angiogenic ECM protein initiates angiogenesis by binding to integrin αvβ5 on resting endothelium, thereby resulting in expression of the transcription factor Hox D3 and integrin αvβ3. Hox D3 converts resting endothelium to angiogenic endothelium by inducing expression of proangiogenic molecules such as integrin αvβ3. These findings provide evidence for an angiogenic switch that can be initiated in the absence of exogenous growth factors and indicate that the angiogenic matrix protein Del-1 may be a useful tool for the therapy of ischemic disease.

Authors

Jingping Zhong, Brian Eliceiri, Dwayne Stupack, Kalyani Penta, Gordon Sakamoto, Thomas Quertermous, Mike Coleman, Nancy Boudreau, Judith A. Varner

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Figure 1

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Del-1 promotes angiogenesis and restores function to murine ischemic mus...
Del-1 promotes angiogenesis and restores function to murine ischemic muscle in vivo. (a) Expression constructs used to induce hDel-1 and hVEGF expression in vivo contained a CMV enhancer/promoter (enh/pro), 5′ UTR, an optimized intron (IVS-8), cDNA encoding either VEGF165 (pVF1164) or full-length human Del-1 (pDL1599), and the hGH 3′ UTR, as well as a kanamycin resistance gene. (b) Expression of Del-1 or VEGF165 in hDel-1 (left) and hVEGF165 (right) transgene-injected, noncoding plasmid–injected, and untreated muscle. Photographs were taken at ×100 magnification. Arrows denote fibers exhibiting positive staining. Cntl, control. (c) Lysates of muscle injected with hDel-1 (Del-1) or noncoding control plasmid as well as purified Del-1 (pur.Del-1) were immunoblotted for Del-1 expression using rabbit anti–Del-1 Ab’s. (d) Mouse model of hind-limb ischemia induced by ligation of the femoral artery. (e and f) Capillary/myofiber ratio 7 days after treatment with noncoding (control), hDel-1, or hVEGF165 expression plasmids. (e) Cryosections of treated muscle immunostained using a rat anti-mouse CD31 Ab. (f) Mean capillary/myofiber ratio ± SEM for three animals per group (*P < 0.01). (g) Hind-limb ischemia was induced by bilateral ligation of the femoral artery of male CD1 mice. Animals were placed on the treadmill for 5 minutes at 5 m/min and then at 10 m/min thereafter. The speed was increased every 2 minutes until fatigue. Results represent the mean ± SEM (n = 7–8 animals/group) treadmill run time for mice treated with formulated noncoding hDel-1 or hVEGF165 expression plasmids. Asterisks indicate statistically significant result relative to noncoding plasmid control (P < 0.05).