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Tissue factor and PAR1 promote microbiota-induced intestinal vascular remodelling

Nature volume 483, pages 627–631 (2012)Cite this article

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Abstract

The gut microbiota is a complex ecosystem that has coevolved with host physiology. Colonization of germ-free (GF) mice with a microbiota promotes increased vessel density in the small intestine1, but little is known about the mechanisms involved. Tissue factor (TF) is the membrane receptor that initiates the extrinsic coagulation pathway2, and it promotes developmental and tumour angiogenesis3,4. Here we show that the gut microbiota promotes TF glycosylation associated with localization of TF on the cell surface, the activation of coagulation proteases, and phosphorylation of the TF cytoplasmic domain in the small intestine. Anti-TF treatment of colonized GF mice decreased microbiota-induced vascular remodelling and expression of the proangiogenic factor angiopoietin-1 (Ang-1) in the small intestine. Mice with a genetic deletion of the TF cytoplasmic domain or with hypomorphic TF (F3) alleles had a decreased intestinal vessel density. Coagulation proteases downstream of TF activate protease-activated receptor (PAR) signalling implicated in angiogenesis5. Vessel density and phosphorylation of the cytoplasmic domain of TF were decreased in small intestine from PAR1-deficient (F2r−/−) but not PAR2-deficient (F2rl1−/−) mice, and inhibition of thrombin showed that thrombin–PAR1 signalling was upstream of TF phosphorylation. Thus, the microbiota-induced extravascular TF–PAR1 signalling loop is a novel pathway that may be modulated to influence vascular remodelling in the small intestine.

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Figure 1: TF promotes microbe-induced vascular remodelling in the gut.
Figure 2: The gut microbiota increases TF procoagulant activity and cell-surface localization.
Figure 3: The gut microbiota increases phosphorylation of the cytoplasmic tail of TF, which increases vessel density in the intestine.
Figure 4: PAR1 activation increases vessel density in the small intestine.

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Acknowledgements

We thank R. Perkins for editing the manuscript; C. Arvidsson, A. Hallén, S. Wagoner, M. Karlsson, D. O’Donell, S. Islam, N. Hörmann and A. Mohammadzadeh for technical assistance; A. Hallén for providing Supplementary Fig. 16; and P. Lindahl, J. Gordon, C. Betsholtz, M. Bergö, A. Wichmann, V. Tremaroli, M. Levin and S. Massberg for comments and suggestions. We are grateful to D. Kirchhofer for the gift of 1H1 monoclonal anti-mouse TF antibody, J. Nichols at Amgen for mL4-3, N. Mackman for the low-TF mice, M. Anderson for the human TF knock-in mice, and J. Gordon for providing CR2-tox176 mice. This study was supported by the Swedish Foundation for Strategic Research, the Swedish Research Council, Torsten and Ragnar Söderberg’s foundation, Petrus and Augusta Hedlund’s foundation, and the Swedish federal government under the LUA/ALF agreement to F.B., National Institutes of Health grants HL-60742 and HL-77753 to W.R., and a Marie Curie Fellowship, a Marie Curie Reintegration Grant from the European Union and the German Federal Ministry of Education and Research (BMBF 01EO1003) to C.R.

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Authors and Affiliations

  1. Sahlgrenska Center for Cardiovascular and Metabolic Research/Wallenberg Laboratory, University of Gothenburg, 413 45 Gothenburg, Sweden,

    Christoph Reinhardt, Mattias Bergentall, Thomas U. Greiner, Gunnel Östergren-Lundén & Fredrik Bäckhed

  2. Department of Molecular and Clinical Medicine, University of Gothenburg, 413 45 Gothenburg, Sweden,

    Christoph Reinhardt, Mattias Bergentall, Thomas U. Greiner, Gunnel Östergren-Lundén & Fredrik Bäckhed

  3. Center for Thrombosis and Hemostasis (CTH), University Medical Center Mainz, Experimental Research, 55131 Mainz, Germany,

    Christoph Reinhardt

  4. Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, 92037, California, USA

    Florence Schaffner & Wolfram Ruf

  5. Haemostasis Biology, Novo Nordisk A/S, DK-2760 Maalov, Denmark,

    Lars C. Petersen

  6. Novo Nordisk Foundation Center for Basic Metabolic Research, Section for Metabolic Receptology and Enteroendocrinology, Faculty of Health Sciences, University of Copenhagen, Copenhagen, DK-2200, Denmark

    Fredrik Bäckhed

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Contributions

C.R. was responsible for conception and study design, biochemical analysis of TF, analysis of vessel densities, data assembly and analysis, and writing the manuscript. M.B., T.U.G., F.S. and G.Ö.L. performed data collection, analysis and interpretation and commented on the manuscript. L.C.P. provided material. W.R. and F.B. were responsible for conception and study design, data analysis and interpretation, and writing the manuscript.

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Correspondence to Fredrik Bäckhed.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-17 and Supplementary Table 1. (PDF 2185 kb)

Supplementary Movie 1

This movie shows a three dimensional rendering of a small intestinal epithelial cell stained for tissue factor (red), cytokeratin (green), and nuclei (blue). (MOV 5 kb)

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Reinhardt, C., Bergentall, M., Greiner, T. et al. Tissue factor and PAR1 promote microbiota-induced intestinal vascular remodelling. Nature 483, 627–631 (2012). https://doi.org/10.1038/nature10893

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