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An antiangiogenic isoform of VEGF-A contributes to impaired vascularization in peripheral artery disease

Nature Medicine volume 20, pages 1464–1471 (2014)Cite this article

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Abstract

Peripheral artery disease (PAD) generates tissue ischemia through arterial occlusions and insufficient collateral vessel formation. Vascular insufficiency in PAD occurs despite higher circulating levels of vascular endothelial growth factor A (VEGF-A)1,2, a key regulator of angiogenesis. Here we show that clinical PAD is associated with elevated levels of an antiangiogenic VEGF-A splice isoform (VEGF-A165b) and a corresponding reduction in levels of the proangiogenic VEGF-A165a splice isoform. In mice, VEGF-A165b expression was upregulated by conditions associated with impaired limb revascularization, including leptin deficiency, diet-induced obesity, genetic ablation of the secreted frizzled-related protein 5 (Sfrp5) adipokine and transgenic overexpression of Wnt5a in myeloid cells. In a mouse model of PAD, delivery of VEGF-A165b inhibited revascularization of ischemic hind limbs, whereas treatment with an isoform-specific neutralizing antibody reversed impaired revascularization caused by metabolic dysfunction or perturbations in the Wnt5a-Sfrp5 regulatory system. These results indicate that inflammation-driven expression of the antiangiogenic VEGF-A isoform can contribute to impaired collateralization in ischemic cardiovascular disease.

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Figure 1: Circulating levels of total VEGF-A, VEGF-A165a and VEGF-A165b in patients with PAD.
Figure 2: Impairment of ischemia-induced angiogenesis by myeloid-specific Wnt5a requires VEGF-A165b.
Figure 3: VEGF-A165b impairs ischemia-induced angiogenesis in mice with diet-induced obesity.
Figure 4: Wnt5a regulates VEGF-A165b expression through a Ror2-JNK-SC35–dependent mechanism in macrophages.

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Acknowledgements

This work was supported by US National Institutes of Health (NIH) grants HL102874, AG34972, HL68758 and HL126141 to K.W. R.K. is supported by the Uehara Memorial Foundation and the Grant-in-Aid for Young Scientists B. N.M.H. is supported by NIH grants HL102299 and HL109790, and N.G. is supported by NIH grants HL081587 and HL1145675. D.O.B. is supported by the British Heart Foundation PG/13/47/30337 and PG/08/054/25272, the Wellcome Trust, Cancer Research UK and the Medical Research Council. We are grateful to S.M. Eswarappa and P.L. Fox, Department of Cellular and Molecular Medicine, The Lerner Research Institute, Cleveland Clinic, for providing Ax-specific antibody and recombinant VEGF-Ax protein.

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Authors and Affiliations

  1. Molecular Cardiology and Whitaker Cardiovascular Institute, Boston University School of Medicine, Boston, Massachusetts, USA

    Ryosuke Kikuchi, Kazuto Nakamura, Susan MacLauchlan, Ippei Shimizu, Jose Javier Fuster, Yasufumi Katanasaka, Sumiko Yoshida, Naomi M Hamburg & Kenneth Walsh

  2. Department of Medical Technique, Nagoya University Hospital, Nagoya, Aichi, Japan

    Ryosuke Kikuchi

  3. Department of Medicine and Whitaker Cardiovascular Institute, Clinical Cardiology, Boston University School of Medicine, Boston, Massachusetts, USA

    Doan Thi-Minh Ngo & Noyan Gokce

  4. Microvascular Research Laboratories, School of Physiology and Pharmacology, University of Bristol, Bristol, UK

    Yan Qiu & David O Bates

  5. Cancer and Developmental Biology Laboratory, National Cancer Institute, National Institutes of Health, Frederick, Maryland, USA

    Terry P Yamaguchi

  6. Department of Clinical Laboratory, Nagoya University Hospital, Nagoya, Aichi, Japan

    Tadashi Matsushita

  7. Department of Cardiology, Nagoya University Graduate School of Medicine, Nagoya, Aichi, Japan

    Toyoaki Murohara

  8. Division of Cancer and Stem Cells, Cancer Biology, School of Medicine, University of Nottingham, Queen's Medical Centre, Nottingham, UK

    David O Bates

  9. Evans Department of Medicine and the Whitaker Cardiovascular Institute, Boston University School of Medicine, Boston, Massachusetts, USA

    Naomi M Hamburg

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  1. Ryosuke Kikuchi
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Contributions

All authors participated in the analysis and interpretation of data. R.K. and N.M.H. performed the statistical analyses of the human and mouse data. R.K. and K.W. conceived the project and designed the experiments. K.N., S.M., D.T.-M.N., I.S., J.J.F., Y.K., S.Y., Y.Q., T. Matsushita, T. Murohara, N.G., D.O.B. and R.K. conducted experiments. R.K., T.P.Y., Y.Q. and D.O.B. developed experimental tools or mouse models. R.K., N.M.H. and K.W. wrote and edited the manuscript.

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Correspondence to Kenneth Walsh.

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The authors declare no competing financial interests.

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Kikuchi, R., Nakamura, K., MacLauchlan, S. et al. An antiangiogenic isoform of VEGF-A contributes to impaired vascularization in peripheral artery disease. Nat Med 20, 1464–1471 (2014). https://doi.org/10.1038/nm.3703

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