ESC Congress 2011 27. 31. August 2011 Paris France VEGFs and the angiogenic paradox in diabetic patients Session: New features and effects of angiogenic growth factors in vascular diseases. Johannes Waltenberger, MD, PhD, F.E.S.C. Professor and Chair of Internal Medicine, Cardiology and Angiology Director, Department of Cardiology and Angiology University Hospital Münster Germany waltenberger@ukmuenster.de
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The angiogenic paradox in diabetes mellitus Definition: The angiogenic paradox describes the fact that diabetes mellitus is associated with both enhanced as well as reduced vascular growth, i.e. angiogenesis (angiogenesis/arteriogenesis). Situation in chronic diabetes mellitus: Diabetic retinopathy enhanced angiogenesis Fewer coronary collaterals reduced arteriogenesis
The angiogenic paradox in diabetes mellitus Physiological response Diabetes mellitus? angiogenesis Diabetic retinopathy arteriogenesis Coronary collaterals
Low VEGF - no vessels - no life Yolk sac of E10.5 VEGF +/+ and VEGF +/ mouse embryos Ferrara. Nature 380(6573):439 442, 1996
Growth factors regulate endothelial function and angiogenesis release from BM VEGF homing anti-thrombosis VEGFR-1 VEGFR-2 anti-apoptosis proliferation migration J. Biol. Chem. 269:26988-95, 1994 J. Biol. Chem. 272:32521-32527, 1997 Biochem. Biophys. Res. Commun. 252:743-6, 1998 Biochem. Biophys. Res. Commun. 265:636-9, 1999 EMBO J. 18:363-374, 1999 J. Clin. Invest. 106:1311-1319, 2000 Biochem. Biophys. Res. Commun. 306: 730-736, 2003 Nat. Med. 9: 936-43, 2003 P Y Y P antiproliferative for SMCs permeability NO PGI 2 Waltenberger, Br. J. Cardiol., 2009
Endothelial Dysfunction release from BM homing anti-thrombosis anti-apoptotisis proliferation migration antiproliferative for SMCs permeability diabetes mellitus hypertension NO PGI 2 hypercholesterolemia smoking Waltenberger, Br. J. Cardiol., 2009
VEGF signalling in endothelial cells and monocytes Endothelial Cell Monocyte PlGF VEGF-A VEGF-E PlGF VEGF-A VEGFR-1 VEGFR-2 Angiogenesis VEGFR-1 P Y Y P P Y Y P P Y Y P Proliferation enos, inos Tissue factor PlGF Angiogenesis Migration NO release Arteriogenesis Endothelial Cells Smooth Muscle Cells Monocytes Tissue factor Migration Waltenberger et al., J. Biol. Chem. 1994 Kroll et al., Biochem. Biophys. Res. Commun. 1998 Kroll et al., Biochem. Biophys. Res. Commun. 1999 Meyer et al., EMBO J. 1999 Autiero*, Waltenberger* et al., Nat. Med. 2003 Clauss et al., J. Biol. Chem. 1996 Waltenberger et al., Circulation 2000 Waltenberger, Cardiovasc. Res. 2005 Stadler et al., Arterioscler. Thromb. Vasc. Biol. 2007 Tchaikovski et al., Arterioscler. Thromb. Vasc. Biol. 2008
The angiogenic paradox in diabetes mellitus Physiological response Diabetes mellitus VEGF level VEGFinduced response angiogenesis Diabetic retinopathy chronic (mo yrs) arteriogenesis Coronary collaterals subacute (wks) Waltenberger J, Biochem Soc Trans. 37: 1167-70, 2009
The angiogenic paradox in diabetes mellitus Angiogenesis Arteriogenesis Vascular Growth Diabetic retinopathy Collateral growth Timing hours / years days to weeks Cell types EC (ectoderm-derived) EC (mesoderm-derived) -- Mo Pathology local VEGF levels (eye) (heart) VEGF response Mo migration - VEGFR-2 levels VEGF signalling EC, endothelial cells; Mo, monocytes
Ligand-induced migration (% of unstimulated control) VEGF-A-induced migration of monocytes is impaired in diabetes mellitus Chemotaxis analysis 300 VEGF-A p < 0.0001 1000 900 fmlp p > 0.1 250 800 monocyte 700 200 600 membrane 500 150 400 ligand 300 100 200 100 50 Non-diabetics Diabetics n = 14 n = 16 0 Non-diabetics Diabetics n = 14 n = 16 Waltenberger, Circulation 102:185-190, 2000
Growth factor dysfunction GF Biochemical modifications of growth factor ligands - oxidation (ROS) - glycation - nitration - other modifications Reduced ligand binding P Y Y P Interference with tyrosine kinase function Impaired signal generation Migration Gene transcription Modifications of signalling molecules - oxidation (ROS) - nitration - other modifications Impaired signal transduction Waltenberger, Cardiovasc. Res. 65: 574-580, 2005
Stimulation of monocytes from diabetics using VEGF-A Intact receptor phosphorylation - Impaired migration PlGF VEGF-A VEGF-A [50 ng/ml] Control Diabetic - + - + Flt-1 VEGFR-1 P Y Y P Diabetes mellitus 200 KDa - 97 KDa - 69 KDa - Migration tissue factor 30 KDa - Arteriogenesis Waltenberger et al., Circulation 102:185-190, 2000
Ligand-induced chemotaxis (% of unstimulated control) VEGFR-1 expression (arbitruary units) VEGFR-1-mediated chemotaxis is reduced in DM, VEGFR-1-mediated while VEGFR-1 expression chemotaxis remains is reduced unchanged in DM, while VEGFR-1 expression remains unchanged 150 p=0.05 7 6 100 5 4 50 3 2 1 0 PlGF-1, 1 ng/ml (n=3) 0 nondm (n=6) DM (n=6) Tchaikovski et al., Circulation,120: 150-159, 2009
VEGFR-1-mediated signalling in monocytes Working hypothesis VEGF-A Flt-1 VEGFR-1 P Y Y P? Migration Arteriogenesis Tchaikovski et al., ATVB 28: 322-328, 2008
Akt-Phosphorylation (compared to non-diabetic Control %) Defective VEGF signalling in diabetic monocytes - Pre-activation of Akt - DM VEGF-A Flt-1 VEGFR-1 400 300 200 p 0.01 p 0.05 p 0.001 Non-Diabetic (17) Diabetic (17) P Y Y P 100 0 P-Akt P-Akt (Ser473) Migration - + - - + - - - + - - + p38 PlGF, 10 ng/ml, 5 VEGF, 10 ng/ml, 5 Arteriogenesis Tchaikovski et al., Circulation,120: 150-159, 2009
PTP1B activity (pmol PO 4 ) Overall PTP activity (pmol PO 4 ) PTPase inhibition promotes monocyte activation and dysfunction in diabetes mellitus Overall PTP activity DM VEGF-A Flt-1 VEGFR-1 400 300 200 100 p<0.05 P Y Y P 0 Non-DM (n=6) DM (n=6) PTPs P-Akt P-ERK1/2 P-p38 Migration Arteriogenesis PTP1B activity 800 700 600 500 400 300 200 100 0 Non-DM (n=6) p=0.055 DM (n=6) Tchaikovski et al., Circulation,120: 150-159, 2009
Defective VEGF signalling in diabetic monocytes - VEGF resistance - AGE PIGF-1 VEGF-A Hyperglycemia RAGE PIGF-1 VEGF-A VEGFR-1?? ROS VEGFR-1 PTPs P-ERK P-p38 P-Akt non-dm PTPs Constitutive elevation in absence of VEGFR-1 stimulation P-ERK P-p38 P-Akt DM Pathways desensitized Tchaikovski et al., Circulation,120: 150-159, 2009
VEGF resistance as the previously missing link to explain the angiogenic paradox Physiological response Diabetes mellitus VEGF level VEGF signaling pathway angiogenesis Diabetic retinopathy chronic (mo yrs) arteriogenesis Coronary collaterals VEGFinduced response subacute (wks) Waltenberger J, Biochem Soc Trans. 37: 1167-70, 2009
The angiogenic paradox in diabetes mellitus Summary: The action of VEGF is essential for endothelial function and angiogenesis. Diabetes mellitus is associated with both enhanced (diabetic retinopathy) as well as reduced vascularization (collateral arteries). Diabetes mellitus is associated with elevated VEGF levels and reduced VEGFR function. VEGF resistance can explain the angiogenic paradox.
Thanks to... Maastricht and Münster Labs on Exp & Molecular Cardiology Nynke van den Akker Frauke S. Czepluch Marjo Donners Marina Dunaeva Mick Gagliardi Fabienne Jeukens Servé Olieslagers Evangelia Pardali Nadina Stadler Nathalie Stonka Vadim Tchaikovski Fons Verheyen Stefan Vöö Münster Collaborators Ralf Adams Frank U. Müller Andreas H. Jakobs Johannes Roth Michael Schäfers Otmar Schober Hans Schöler Lydia Sorokin Dietmar Vestweber External Collaborators Frank-D Böhmer Kenneth Caidahl Hugo ten Cate Peter Carmeliet Lena Claesson-Welsh Mat Daemen Peter ten Dijke Carl-Henrik Heldin Jens Kroll Tim Leiner Charles Lapière Karl Plate Mark J. Post Frits Prinzen Masabumi Shibuya Marina Ziche Juleen Zierath INTACT 1