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Macrophage skewing by Phd2 haplodeficiency prevents ischaemia by inducing arteriogenesis

Nature volume 479, pages 122–126 (2011)Cite this article

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Abstract

PHD2 serves as an oxygen sensor that rescues blood supply by regulating vessel formation and shape in case of oxygen shortage1,2,3,4,5. However, it is unknown whether PHD2 can influence arteriogenesis. Here we studied the role of PHD2 in collateral artery growth by using hindlimb ischaemia as a model, a process that compensates for the lack of blood flow in case of major arterial occlusion6,7,8. We show that Phd2 (also known as Egln1) haplodeficient (Phd2+/−) mice displayed preformed collateral arteries that preserved limb perfusion and prevented tissue necrosis in ischaemia. Improved arteriogenesis in Phd2+/− mice was due to an expansion of tissue-resident, M2-like macrophages9,10 and their increased release of arteriogenic factors, leading to enhanced smooth muscle cell (SMC) recruitment and growth. Both chronic and acute deletion of one Phd2 allele in macrophages was sufficient to skew their polarization towards a pro-arteriogenic phenotype. Mechanistically, collateral vessel preconditioning relied on the activation of canonical NF-κB pathway in Phd2+/− macrophages. These results unravel how PHD2 regulates arteriogenesis and artery homeostasis by controlling a specific differentiation state in macrophages and suggest new treatment options for ischaemic disorders.

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Figure 1: Phd2 +/− mice are pre-adapted to ischaemia.
Figure 2: Phd2 +/− macrophages display an M2-like phenotype.
Figure 3: Phd2 +/− macrophages protect against ischaemia by inducing arteriogenesis.
Figure 4: Phd2 +/− macrophages trigger arteriogenesis via canonical NF-κB pathway.

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Acknowledgements

This work was supported by grants from FWO (G.0726.10), Belgium, and from VIB. The authors are thankful to P. Carmeliet for the Phd2 KO and cKO mice, M. Karin for the Ikbkb cKO mice, P. Ratcliffe for the PHD2H313A construct, A. Luttun and P. Fazzari for comments, Y. Jonsson and T. Janssens for technical assistance. VE-Cadherin:CreERT and PDGFRB:Cre transgenic mice were generated at Cancer Research UK and kindly donated by R. Adams. E.D. was funded by ARC, S.C. by FCT, R.L.O. and V.F. by FWO, A.H. by DFG. C.R. was supported by COST action TD0901. M.D.P. was supported by an ERC starting grant.

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Author notes

  1. Tapan Bhattacharyya & Michele De Palma

    Present address: Present addresses: Faculty of Infectious & Tropical Diseases, London School of Hygiene & Tropical Medicine, London WC1E 7HT, UK (T.P.); The Swiss Institute for Experimental Cancer Research (ISREC), Swiss Federal Institute of Technology Lausanne (EPFL), Lausanne, CH-1015, Switzerland (M.D.P.).,

  2. Sandra Costa, Estelle Delamarre and Carmen Roncal: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Molecular Oncology and Angiogenesis, Vesalius Research Center, VIB, Leuven B-3000, Belgium

    Yukiji Takeda, Sandra Costa, Estelle Delamarre, Carmen Roncal, Rodrigo Leite de Oliveira, Veronica Finisguerra, Sofie Deschoemaeker, Mathias Wenes, Alexander Hamm, Jens Serneels & Massimiliano Mazzone

  2. Laboratory of Molecular Oncology and Angiogenesis, Vesalius Research Center, K. U. Leuven, Leuven B-3000, Belgium

    Yukiji Takeda, Sandra Costa, Estelle Delamarre, Carmen Roncal, Rodrigo Leite de Oliveira, Veronica Finisguerra, Sofie Deschoemaeker, Mathias Wenes, Alexander Hamm, Jens Serneels & Massimiliano Mazzone

  3. Laboratory of Angiogenesis and Neurovascular link, Vesalius Research Center, VIB, Leuven B-3000, Belgium

    Yukiji Takeda, Carmen Roncal, Rodrigo Leite de Oliveira & Françoise Bruyère

  4. Laboratory of Angiogenesis and Neurovascular link, Vesalius Research Center, K. U. Leuven, Leuven B-3000, Belgium

    Yukiji Takeda, Carmen Roncal, Rodrigo Leite de Oliveira & Françoise Bruyère

  5. Life and Health Sciences Research Institute, Minho University, 4710-057 Braga, Portugal

    Sandra Costa

  6. Atherosclerosis Research Laboratory, CIMA-University of Navarra, 31008 Pamplona, Spain

    Carmen Roncal

  7. Angiogenesis and Tumor Targeting Unit and HSR-TIGET, San Raffaele Institute, 20132 Milan, Italy

    Mario Leonardo Squadrito & Michele De Palma

  8. Vita-Salute University, 20132 Milan, Italy

    Mario Leonardo Squadrito

  9. Biomedical Magnetic Resonance Unit, Universite Catholique de Louvain, Brussels B-1200, Belgium

    Julie Magat, Benedicte F. Jordan & Bernard Gallez

  10. Rayne Institute, University College London, London WC1E 6JF, UK

    Tapan Bhattacharyya & Patrick Maxwell

  11. Molecular/Cancer Biology Laboratory, Research Programs Unit and Institute for Molecular Medicine, Biomedicum Helsinki, 00014 Helsinki, Finland

    Andrey Anisimov & Kari Alitalo

  12. Cardiovascular Medicine, Yale University School of Medicine, New Haven, 06510, Connecticut, USA

    Zhen W. Zhuang & Michael Simons

  13. The First Department of Internal Medicine, Nara Medical University, 634-8522 Nara, Japan

    Yoshihiko Saito

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Contributions

Y.T., E.D. and S.C. performed experimental design, all experiments, acquisition of data and analysis and interpretation of all data. C.R. performed analysis of histological stainings, angiographies. R.L.O., C.R. and S.C. performed the western blots. R.L.O. and V.F. performed treadmill-running tests, quantitative PCR experiments and drug administrations. M.L.S. performed lentiviral vector preparation and cell transduction. F.B. performed EC isolation and angiography measurements. J.M., B.F.J. and B.G. performed oxymetry experiments. S.D. performed luciferase assays. M.W. and A.H. performed transplantation experiments and electroporations. Y.T. and J.S. performed the ligations of the femoral artery. Z.W.Z. and M.S. performed micro-computer tomography angiograms. A.A. and K.A. contributed vital reagents. T.B. and P.M. contributed in generating the Phd2 targeting vector. Y.T., E.D., S.C., C.R., Y.S. and M.D.P. participated in scientific discussion and drafting of the manuscript. M.M. performed experimental design, data analysis, conducted scientific direction and wrote the manuscript.

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Correspondence to Massimiliano Mazzone.

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The authors declare no competing financial interests.

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Takeda, Y., Costa, S., Delamarre, E. et al. Macrophage skewing by Phd2 haplodeficiency prevents ischaemia by inducing arteriogenesis. Nature 479, 122–126 (2011). https://doi.org/10.1038/nature10507

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