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Endoglin prevents vascular malformation by regulating flow-induced cell migration and specification through VEGFR2 signalling

Nature Cell Biology volume 19, pages 639652 (2017) | Download Citation


Loss-of-function (LOF) mutations in the endothelial cell (EC)-enriched gene endoglin (ENG) cause the human disease hereditary haemorrhagic telangiectasia-1, characterized by vascular malformations promoted by vascular endothelial growth factor A (VEGFA). How ENG deficiency alters EC behaviour to trigger these anomalies is not understood. Mosaic ENG deletion in the postnatal mouse rendered Eng LOF ECs insensitive to flow-mediated venous to arterial migration. Eng LOF ECs retained within arterioles acquired venous characteristics and secondary ENG-independent proliferation resulting in arteriovenous malformation (AVM). Analysis following simultaneous Eng LOF and overexpression (OE) revealed that ENG OE ECs dominate tip-cell positions and home preferentially to arteries. ENG knockdown altered VEGFA-mediated VEGFR2 kinetics and promoted AKT signalling. Blockage of PI(3)K/AKT partly normalized flow-directed migration of ENG LOF ECs in vitro and reduced the severity of AVM in vivo. This demonstrates the requirement of ENG in flow-mediated migration and modulation of VEGFR2 signalling in vascular patterning.

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We thank R. Adams (Max Planck Institute for Molecular Biomedicine, Germany) for providing the Cdh5(PAC)-CreERT2 mice, C. Vary (Michigan State University, USA) for the ENGiOE mice, G. Thurston (Regeneron, USA) for providing the EngLacZ mice, and B. Lavina and K. Gaengel (Uppsala University, Sweden) for sharing mice. We are grateful to Y. Xiong for help on the construction and package of lentivirus and to A. Keller, M. A. Mäe and K. Pietras for initial contributions. This study was supported by grants from William K. Bowes Jr Foundation (L.J.), the Swedish Research Council (L.J., C.B.), the Swedish Cancer Society (L.J., C.B.), the Cardiovascular Programme and the Strategic Research Programme in Neuroscience at Karolinska Institutet (L.J.), Jeanssons Stiftelser (L.J.), Magnus Bergvalls Stiftelse (L.J.), Knut and Alice Wallenbergs Stiftelse (C.B.), the European Research Council (C.B.), the Leducq Foundation (C.B.) and the British Heart Foundation (H.M.A.; RG/12/2/29416).

Author information


  1. Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Scheeles väg 2, 171 77 Stockholm, Sweden

    • Yi Jin
    • , Lars Muhl
    • , Mikhail Burmakin
    • , Yixin Wang
    • , Anne-Claire Duchez
    •  & Lars Jakobsson
  2. Department of Immunology, Genetics and Pathology, Uppsala University, Dag Hammarskjölds väg 20, 751 85 Uppsala, Sweden

    • Christer Betsholtz
  3. Integrated Cardio Metabolic Centre (ICMC), Karolinska Institutet, Novum, Blickagången 6, SE14157 Huddinge, Sweden

    • Christer Betsholtz
  4. Institute of Genetic Medicine, International Centre for Life, Newcastle University, Newcastle upon Tyne NE1 3BZ, UK

    • Helen M. Arthur


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L.J., Y.J. and L.M. designed the research; Y.J., L.M., C.B., H.M.A. and L.J. wrote the paper; Y.J., L.M., Y.W., M.B. and A.-C.D. performed the experiments and together with L.J. analysed the data. All authors discussed the results and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Lars Jakobsson.

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  1. 1.

    3D-visualization of individual EC behaviour in the mouse aortic ring assay.

    See Methods for the setup for imaging. Images were processed in Imaris for surface rendering of mCherry-labelled endothelial nuclei (green). Migratory routes of the ECs are shown with time-indicating tracks. Note the differences in the migratory speed and direction of individual ECs at different time.

  2. 2.

    Migration of control HDMECs under flow.

    See Methods for details on microscopy. Coloured dots and tracks are the migratory routes of all the trackable cells in a randomly chosen field.

  3. 3.

    Migration of ENG knockdown HDMECs under flow.

    See Methods for details on microscopy. Coloured dots and tracks are the migratory routes of all the trackable cells in a randomly chosen field.

  4. 4.

    Analysis of EC migration in vessels of the wounded mouse cornea, by intravital imaging.

    In vivo imaging of newly formed vessels in the mouse cornea 5 days after suturing. YFP expression (green) was induced in a subpopulation of ECs. Blood flow was visualized by transmitted light (100 ms/frame). Arrows indicate leukocytes rolling with the blood flow.

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