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The succinate receptor GPR91 in neurons has a major role in retinal angiogenesis

Nature Medicine volume 14, pages 1067–1076 (2008)Cite this article

Abstract

Vascularization is essential for tissue development and in restoration of tissue integrity after an ischemic injury. In studies of vascularization, the focus has largely been placed on vascular endothelial growth factor (VEGF), yet other factors may also orchestrate this process. Here we show that succinate accumulates in the hypoxic retina of rodents and, via its cognate receptor G protein–coupled receptor-91 (GPR91), is a potent mediator of vessel growth in the settings of both normal retinal development and proliferative ischemic retinopathy. The effects of GPR91 are mediated by retinal ganglion neurons (RGCs), which, in response to increased succinate levels, regulate the production of numerous angiogenic factors including VEGF. Accordingly, succinate did not have proangiogenic effects in RGC-deficient rats. Our observations show a pathway of metabolite signaling where succinate, acting through GPR91, governs retinal angiogenesis and show the propensity of RGCs to act as sensors of ischemic stress. These findings provide a new therapeutic target for modulating revascularization.

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Figure 1: Succinate levels rise in the ischemic retina, and GPR91 is strongly and predominantly expressed by retinal ganglion neurons.
Figure 2: Developmental retinal vessel growth is influenced by succinate and GPR91.
Figure 3: Retinal ganglion neurons mediate the proangiogenic effects of succinate via GPR91.
Figure 4: Succinate stimulates the production of proangiogenic factors by RGCs via a HIF-1α-independent pathway.
Figure 5: Retinal ganglion neuron ablation impedes succinate-induced neovascularization in the adult.
Figure 6: GPR91 knockdown protects against retinal neovascularization in ischemic proliferative retinopathy.

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References

  1. Adair, T.H., Gay, W.J. & Montani, J.P. Growth regulation of the vascular system: evidence for a metabolic hypothesis. Am. J. Physiol. 259, R393–R404 (1990).

    PubMed  CAS  Google Scholar 

  2. Aiello, L.P. Vascular endothelial growth factor and the eye: biochemical mechanisms of action and implications for novel therapies. Ophthalmic Res. 29, 354–362 (1997).

    Article  PubMed  CAS  Google Scholar 

  3. Enge, M. et al. Endothelium-specific platelet-derived growth factor-B ablation mimics diabetic retinopathy. EMBO J. 21, 4307–4316 (2002).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  4. Smith, L.E. et al. Essential role of growth hormone in ischemia-induced retinal neovascularization. Science 276, 1706–1709 (1997).

    Article  PubMed  CAS  Google Scholar 

  5. Watanabe, D. et al. Erythropoietin as a retinal angiogenic factor in proliferative diabetic retinopathy. N. Engl. J. Med. 353, 782–792 (2005).

    Article  PubMed  CAS  Google Scholar 

  6. D'Amore, P.A. Mechanisms of retinal and choroidal neovascularization. Invest. Ophthalmol. Vis. Sci. 35, 3974–3979 (1994).

    PubMed  CAS  Google Scholar 

  7. Cai, W., Rook, S.L., Jiang, Z.Y., Takahara, N. & Aiello, L.P. Mechanisms of hepatocyte growth factor–induced retinal endothelial cell migration and growth. Invest. Ophthalmol. Vis. Sci. 41, 1885–1893 (2000).

    PubMed  CAS  Google Scholar 

  8. Folbergrova, J., Ljunggren, B., Norberg, K. & Siesjo, B.K. Influence of complete ischemia on glycolytic metabolites, citric acid cycle intermediates, and associated amino acids in the rat cerebral cortex. Brain Res. 80, 265–279 (1974).

    Article  PubMed  CAS  Google Scholar 

  9. Hoyer, S. & Krier, C. Ischemia and aging brain. Studies on glucose and energy metabolism in rat cerebral cortex. Neurobiol. Aging 7, 23–29 (1986).

    Article  PubMed  CAS  Google Scholar 

  10. Gutman, M., Bonomi, F., Pagani, S., Cerletti, P. & Kroneck, P. Modulation of the flavin redox potential as mode of regulation of succinate dehydrogenase activity. Biochim. Biophys. Acta 591, 400–408 (1980).

    Article  PubMed  CAS  Google Scholar 

  11. Meixner-Monori, B., Kubicek, C.P., Habison, A., Kubicek-Pranz, E.M. & Rohr, M. Presence and regulation of the α-ketoglutarate dehydrogenase multienzyme complex in the filamentous fungus Aspergillus niger. J. Bacteriol. 161, 265–271 (1985).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  12. Mailloux, R.J. et al. The tricarboxylic acid cycle, an ancient metabolic network with a novel twist. PLoS ONE 2, e690 (2007).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  13. Burns, P.A. & Wilson, D.J. Angiogenesis mediated by metabolites is dependent on vascular endothelial growth factor (VEGF). Angiogenesis 6, 73–77 (2003).

    Article  PubMed  CAS  Google Scholar 

  14. Lee, M.S. et al. Angiogenic activity of pyruvic acid in in vivo and in vitro angiogenesis models. Cancer Res. 61, 3290–3293 (2001).

    PubMed  CAS  Google Scholar 

  15. Murray, B. & Wilson, D.J. A study of metabolites as intermediate effectors in angiogenesis. Angiogenesis 4, 71–77 (2001).

    Article  PubMed  CAS  Google Scholar 

  16. Neuman, R.E. & Mc, C.T. Growth-promoting properties of pyruvate oxal-acetate, and α-ketoglutarate for isolated Walker carcinosarcoma 256 cells. Proc. Soc. Exp. Biol. Med. 98, 303–306 (1958).

    Article  PubMed  CAS  Google Scholar 

  17. He, W. et al. Citric acid cycle intermediates as ligands for orphan G protein–coupled receptors. Nature 429, 188–193 (2004).

    Article  PubMed  CAS  Google Scholar 

  18. Sennlaub, F. et al. Cyclooxygenase-2 in human and experimental ischemic proliferative retinopathy. Circulation 108, 198–204 (2003).

    Article  PubMed  CAS  Google Scholar 

  19. Kushnir, M.M., Komaromy-Hiller, G., Shushan, B., Urry, F.M. & Roberts, W.L. Analysis of dicarboxylic acids by tandem mass spectrometry. High-throughput quantitative measurement of methylmalonic acid in serum, plasma, and urine. Clin. Chem. 47, 1993–2002 (2001).

    Article  PubMed  CAS  Google Scholar 

  20. van Adel, B.A., Kostic, C., Deglon, N., Ball, A.K. & Arsenijevic, Y. Delivery of ciliary neurotrophic factor via lentiviral-mediated transfer protects axotomized retinal ganglion cells for an extended period of time. Hum. Gene Ther. 14, 103–115 (2003).

    Article  PubMed  CAS  Google Scholar 

  21. Wittenberger, T. et al. GPR99, a new G protein–coupled receptor with homology to a new subgroup of nucleotide receptors. BMC Genomics 3, 17 (2002).

    Article  PubMed  PubMed Central  Google Scholar 

  22. Ogunshola, O.O. et al. Neuronal VEGF expression correlates with angiogenesis in postnatal developing rat brain. Brain Res. Dev. Brain. Res. 119, 139–153 (2000).

    Article  PubMed  CAS  Google Scholar 

  23. Marti, H.H. & Risau, W. Systemic hypoxia changes the organ-specific distribution of vascular endothelial growth factor and its receptors. Proc. Natl. Acad. Sci. USA 95, 15809–15814 (1998).

    Article  PubMed  CAS  Google Scholar 

  24. Stowe, A.M. et al. VEGF protein associates to neurons in remote regions following cortical infarct. J. Cereb. Blood Flow Metab. 27, 76–85 (2007).

    Article  PubMed  CAS  Google Scholar 

  25. Calza, L., Giardino, L., Giuliani, A., Aloe, L. & Levi-Montalcini, R. Nerve growth factor control of neuronal expression of angiogenetic and vasoactive factors. Proc. Natl. Acad. Sci. USA 98, 4160–4165 (2001).

    Article  PubMed  CAS  Google Scholar 

  26. Iriyama, A., Chen, Y.N., Tamaki, Y. & Yanagi, Y. Effect of anti-VEGF antibody on retinal ganglion cells in rats. Br. J. Ophthalmol. 91, 1230–1233 (2007).

    Article  PubMed  PubMed Central  Google Scholar 

  27. Zhu, T. et al. Proangiogenic effects of protease-activated receptor 2 are tumor necrosis factor-α and consecutively Tie2 dependent. Arterioscler. Thromb. Vasc. Biol. 26, 744–750 (2006).

    Article  PubMed  CAS  Google Scholar 

  28. Eklund, L. & Olsen, B.R. Tie receptors and their angiopoietin ligands are context-dependent regulators of vascular remodeling. Exp. Cell Res. 312, 630–641 (2006).

    Article  PubMed  CAS  Google Scholar 

  29. Koivunen, P. et al. Inhibition of hypoxia-inducible factor (HIF) hydroxylases by citric acid cycle intermediates: possible links between cell metabolism and stabilization of HIF. J. Biol. Chem. 282, 4524–4532 (2007).

    Article  PubMed  CAS  Google Scholar 

  30. Selak, M.A. et al. Succinate links TCA cycle dysfunction to oncogenesis by inhibiting HIF-α prolyl hydroxylase. Cancer Cell 7, 77–85 (2005).

    Article  PubMed  CAS  Google Scholar 

  31. McColm, J.R., Geisen, P. & Hartnett, M.E. VEGF isoforms and their expression after a single episode of hypoxia or repeated fluctuations between hyperoxia and hypoxia: relevance to clinical ROP. Mol. Vis. 10, 512–520 (2004).

    PubMed  PubMed Central  CAS  Google Scholar 

  32. Ikeda, M., Hosoda, Y., Hirose, S., Okada, Y. & Ikeda, E. Expression of vascular endothelial growth factor isoforms and their receptors Flt-1, KDR and neuropilin-1 in synovial tissues of rheumatoid arthritis. J. Pathol. 191, 426–433 (2000).

    Article  PubMed  CAS  Google Scholar 

  33. Usui, T. et al. VEGF164(165) as the pathological isoform: differential leukocyte and endothelial responses through VEGFR1 and VEGFR2. Invest. Ophthalmol. Vis. Sci. 45, 368–374 (2004).

    Article  PubMed  Google Scholar 

  34. Sapieha, P.S., Peltier, M., Rendahl, K.G., Manning, W.C. & Di Polo, A. Fibroblast growth factor-2 gene delivery stimulates axon growth by adult retinal ganglion cells after acute optic nerve injury. Mol. Cell. Neurosci. 24, 656–672 (2003).

    Article  PubMed  CAS  Google Scholar 

  35. Berkelaar, M., Clarke, D.B., Wang, Y.C., Bray, G.M. & Aguayo, A.J. Axotomy results in delayed death and apoptosis of retinal ganglion cells in adult rats. J. Neurosci. 14, 4368–4374 (1994).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  36. Huxlin, K.R., Dreher, Z., Schulz, M. & Dreher, B. Glial reactivity in the retina of adult rats. Glia 15, 105–118 (1995).

    Article  PubMed  CAS  Google Scholar 

  37. Mu, X. et al. Ganglion cells are required for normal progenitor cell proliferation but not cell-fate determination or patterning in the developing mouse retina. Curr. Biol. 15, 525–530 (2005).

    Article  PubMed  CAS  Google Scholar 

  38. Higgins, R.D. et al. Diltiazem reduces retinal neovascularization in a mouse model of oxygen induced retinopathy. Curr. Eye Res. 18, 20–27 (1999).

    Article  PubMed  CAS  Google Scholar 

  39. Gariano, R.F. & Gardner, T.W. Retinal angiogenesis in development and disease. Nature 438, 960–966 (2005).

    Article  PubMed  CAS  Google Scholar 

  40. Arjamaa, O. & Nikinmaa, M. Oxygen-dependent diseases in the retina: role of hypoxia-inducible factors. Exp. Eye Res. 83, 473–483 (2006).

    Article  PubMed  CAS  Google Scholar 

  41. Shweiki, D., Itin, A., Soffer, D. & Keshet, E. Vascular endothelial growth factor induced by hypoxia may mediate hypoxia-initiated angiogenesis. Nature 359, 843–845 (1992).

    Article  CAS  PubMed  Google Scholar 

  42. Johnson, R.N. & Hansford, R.G. The control of tricarboxylate-cycle oxidations in blowfly flight muscle. The steady-state concentrations of citrate, isocitrate 2-oxoglutarate and malate in flight muscle and isolated mitochondria. Biochem. J. 146, 527–535 (1975).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  43. Hems, D.A. & Brosnan, J.T. Effects of ischaemia on content of metabolites in rat liver and kidney in vivo. Biochem. J. 120, 105–111 (1970).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  44. Pierce, E.A., Avery, R.L., Foley, E.D., Aiello, L.P. & Smith, L.E. Vascular endothelial growth factor/vascular permeability factor expression in a mouse model of retinal neovascularization. Proc. Natl. Acad. Sci. USA 92, 905–909 (1995).

    Article  PubMed  CAS  Google Scholar 

  45. Smith, L.E. et al. Oxygen-induced retinopathy in the mouse. Invest. Ophthalmol. Vis. Sci. 35, 101–111 (1994).

    PubMed  CAS  Google Scholar 

  46. Matthews, P.M., Nagy, Z., Brown, G.K., Land, J. & Squier, M.V. Isolated capillary proliferation in Leigh's syndrome. Clin. Neuropathol. 13, 139–141 (1994).

    PubMed  CAS  Google Scholar 

  47. Piao, Y.S., Tang, G.C., Yang, H. & Lu, D.H. Clinico-neuropathological study of a Chinese case of familial adult Leigh syndrome. Neuropathology 26, 218–221 (2006).

    Article  PubMed  Google Scholar 

  48. Francois, J. & Neetens, A. Comparative anatomy of the vascular supply of the eye in vertebrates. in In The Eye Vol. 5, (ed. H. Davson & L.T. Graham) 1–70 (Academic Press, New York, 1974).

    Google Scholar 

  49. Johnson, G.L. Ophthalmoscopic studies on the eyes of mammals. Phil. Trans. R. Soc. Lond. B2, 1–82 (1968).

    Google Scholar 

  50. Michaelson, I.C. Retinal Circulation in Man and Mammals. (ed. Thomas, C.C.) Ch. 1–9 (Charles C. Thomas, Springfield, Illinois, 1954).

    Google Scholar 

  51. Braun, R.D., Linsenmeier, R.A. & Goldstick, T.K. Oxygen consumption in the inner and outer retina of the cat. Invest. Ophthalmol. Vis. Sci. 36, 542–554 (1995).

    PubMed  CAS  Google Scholar 

  52. Fruttiger, M. et al. PDGF mediates a neuron-astrocyte interaction in the developing retina. Neuron 17, 1117–1131 (1996).

    Article  PubMed  CAS  Google Scholar 

  53. Hughes, S., Yang, H. & Chan-Ling, T. Vascularization of the human fetal retina: roles of vasculogenesis and angiogenesis. Invest. Ophthalmol. Vis. Sci. 41, 1217–1228 (2000).

    PubMed  CAS  Google Scholar 

  54. Dreher, B. & Robinson, S.R. Development of the retinofugal pathway in birds and mammals: evidence for a common 'timetable'. Brain Behav. Evol. 31, 369–390 (1988).

    Article  PubMed  CAS  Google Scholar 

  55. Cringle, S.J., Yu, P.K., Su, E.N. & Yu, D.Y. Oxygen distribution and consumption in the developing rat retina. Invest. Ophthalmol. Vis. Sci. 47, 4072–4076 (2006).

    Article  PubMed  Google Scholar 

  56. Arany, Z. et al. HIF-independent regulation of VEGF and angiogenesis by the transcriptional coactivator PGC-1α. Nature 451, 1008–1012 (2008).

    Article  PubMed  CAS  Google Scholar 

  57. Frassetto, L.J. et al. Kinase-dependent differentiation of a retinal ganglion cell precursor. Invest. Ophthalmol. Vis. Sci. 47, 427–438 (2006).

    Article  PubMed  Google Scholar 

  58. Dull, T. et al. A third-generation lentivirus vector with a conditional packaging system. J. Virol. 72, 8463–8471 (1998).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  59. Penn, J.S., Henry, M.M. & Tolman, B.L. Exposure to alternating hypoxia and hyperoxia causes severe proliferative retinopathy in the newborn rat. Pediatr. Res. 36, 724–731 (1994).

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the Canadian Institutes of Health Research, the March of Dimes Birth Defects Foundation, the Heart and Stroke Foundation of Québec and the Fonds de la Recherche en Santé du Québec. P.S. and M.S. hold a Research Fellowship Award and a studentship from the Heart and Stroke Foundation of Canada, respectively. K.Z. is a recipient of The Foundation Fighting Blindness Postdoctoral Fellowship Award. F.S. and S.C. are recipients of a fellowship and scientist awards, respectively, from the Canadian Institutes of Health Research. S.C. also holds a Canada Research Chair (perinatology). Generation of brn3bZ-dta/+;six3-cre mice and J.-H.C.'s salary were supported by a US National Eye Institute grant EY011930 to W.H.K. and by the Robert A. Welch Foundation (G-0010), respectively. We wish to thank N. Agarwal, from the University of North Texas Health Science Center at Fort Worth, for his kind donation of the RGC-5 cell line. We also wish to thank H. Fernandez and S. Leclerc for valuable technical assistance.

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  1. Przemyslaw Sapieha and Mirna Sirinyan: These authors contributed equally to this work.

Authors and Affiliations

  1. Departments of Pediatrics and Pharmacology, Research Center of Centre Hospitalier Universitaire Sainte-Justine, Université de Montréal, Montreal, H3T 1C5, Quebec, Canada

    Przemyslaw Sapieha, Mirna Sirinyan, David Hamel, Karine Zaniolo, Jean-Sébastien Joyal, Jean-Claude Honoré, Sophie Tremblay, Martin Leduc, Lenka Rihakova, Pierre Hardy, Christian Beauséjour, Gregor Andelfinger, Grant Mitchell & Sylvain Chemtob

  2. Department of Pharmacology and Therapeutics, McGill University, Montreal, H3G 1Y6, Quebec, Canada

    Przemyslaw Sapieha, Mirna Sirinyan, Jean-Sébastien Joyal & Daya R Varma

  3. Department of Biochemistry and Molecular Biology, University of Texas M.D. Anderson Cancer Center, Houston, 77030, Texas, USA

    Jang-Hyeon Cho, William H Klein & Xiuqian Mu

  4. Institut National de la Santé et de la Recherche Médicale, Unité Mixte de Recherche Scientique S 872, Centre de Recherche des Cordeliers, Paris, F-75006, France

    Elsa Kermorvant-Duchemin & Florian Sennlaub

  5. Department of Medicine, Mass Spectrometry Facility, McGill University Montreal, H3A 1A4, Quebec, Canada

    Orval Mamer

  6. Departments of Ophthalmology and Neurology-Neurosurgery, McGill University-Montreal Children's Hospital Research Institute, Montreal, H3H 1P3, Quebec, Canada

    Pierre Lachapelle

  7. Department of Pathology and Cell Biology, University of Montreal, Montreal, H3T 1J4, Quebec, Canada

    Adriana Di Polo

  8. Université Pierre et Marie Curie (Paris 6), Unité Mixte de Recherche Scientique S 872, Paris, F-75006, France

    Florian Sennlaub

  9. Université Paris Descartes, Unité Mixte de Recherche Scientique 872, Paris, F-75006, France

    Florian Sennlaub

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Correspondence to Sylvain Chemtob.

Supplementary information

Supplementary Text and Figures

Supplementary Figs. 1-4, Supplementary Table 1 and Supplementary Methods (PDF 1751 kb)

Supplementary Video 1

Three-dimensional confocal representation of a 30-μm–thick radial section of the retina. GPR91-positive cells are labeled in green, whereas lectin-stained blood vessels are in red. A lack of overlap indicates that GPR91 is not expressed in retinal blood vessels. (MOV 1060 kb)

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Sapieha, P., Sirinyan, M., Hamel, D. et al. The succinate receptor GPR91 in neurons has a major role in retinal angiogenesis. Nat Med 14, 1067–1076 (2008). https://doi.org/10.1038/nm.1873

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