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1 DECLARATION OF CONFLICT OF INTEREST No conflicts of interest
2 University Heart Centre Tübingen Angiogenic actions of platelets Meinrad Gawaz, MD, FESC Tübingen, Germany ESC 2011 Paris
3 GPIb GPIb GPVI TxA2 ADP GPIb GPIIb/IIIa GPVI
4 Platelet Interaction with Vascular Cells Recruitment of bone marrow-derived cells (BMDC) Angiogenesis Proteolysis upa/upar, EMMPRIN MMPs Thrombosis Tissue Factor Cell Differentiation to Macrophages & Foam cells Adhesion Migration BMDC Adhesion receptors VCAM-1, ICAM-1, avß3, E-selectin Proteolysis upa/upar, MMPs Platelet NF-kB Monocyte adhesion & secretion adhesion & secretion Endothelial cell Chemotaxis MCP-1, MIP1-a Adhesion Thrombosis Inflammation TNF-a, IL-8 Mac-1,VLA-4, CD62L PMN Cytokines & cytokine-like factors MCP-1, CD40L Tissue Factor, PAI-1 Oxidative burst Proteolysis upa/upa-r, MMPs, Cathepsin-G Endothelial cell activation and migration JCI 2005; ATVB 2008
5 Angiogenesis Angiogenic Factors 1. Lysis of ECM and base membrane 2. EC Invasion/ Migration/ Proliferation 3. Capillary tube Formation and Differentiation Angiogenesis Tube Formation Assay (modified according BD Biosciences)
6 Angiogenesis Platelets? 1. Lysis of ECM and base membrane 2. EC Invasion/ Migration/ Proliferation 3. Capillary tube Formation and Differentiation Angiogenesis Tube Formation Assay (modified according BD Biosciences)
7 Platelets Store and Release Angiogenic Factors Platelet Platelet Endothel Endothel Resting Activated Stellos et al. Sem Thromb Haemost 2010
8 Platelet Interaction with Vascular Cells Recruitment of bone marrow-derived cells (BMDC) Angiogenesis Proteolysis upa/upar, EMMPRIN MMPs Thrombosis Tissue Factor Cell Differentiation to Macrophages & Foam cells Adhesion Migration BMDC Adhesion receptors VCAM-1, ICAM-1, avß3, E-selectin Proteolysis upa/upar, MMPs Platelet NF-kB Monocyte adhesion & secretion adhesion & secretion Endothelial cell Chemotaxis MCP-1, MIP1-a Adhesion Thrombosis Inflammation TNF-a, IL-8 Mac-1,VLA-4, CD62L PMN Cytokines & cytokine-like factors MCP-1, CD40L Tissue Factor, PAI-1 Oxidative burst Proteolysis upa/upa-r, MMPs, Cathepsin-G Endothelial cell activation and migration JCI 2005; ATVB 2008
9 Platelets Adhere to and Activate Endothelial Cells Confocal laser microscopy Platelet adhesion to activated endothelial cells 40 (%) 30 CD41 immunofluorescence control GPIIb-IIIa Inhibitor Gawaz et al., Circulation 1996, 1997
10 Platelets Induce Endothelial Inflammation, Migration & ECM-Degradation Platelets Adhesion VCAM-1 MCP-1 Migration Chemotaxis IKK-a/b NF-kB IkB IkB p65 p50 p38 Nucleus upar MMP Proteolysis Gawaz M. Cardiovasc Res, 2004
11 Platelets contain significant amounts of pro- (e.g. VEGF) and anti-angiogenic (e.g. endostatin) factors. VEGF Endostatin Italiano et al. Blood 2009
12 ADP mediates release of VEGF from platelets by American Society of Hematology Battinelli E M et al. Blood 2011;118:
13 Promotion of angiogenesis by ADP-stimulated platelet releasates. Battinelli E M et al. Blood 2011;118: by American Society of Hematology
14 PAR1 and PAR4 agonists differential release of VEGF and endostatin
15 Platelet levels alter blood flow in ischemic limbs. Feng W et al. Blood 2011;117: by American Society of Hematology
16 Blood vessel density is diminished on platelet depletion. Feng W et al. Blood 2011;117: by American Society of Hematology
17 Platelet microparticles (PMP) induce angiogenesis in a dose dependent fashion Brill A et al. Cardiovasc Res 2005, 67: 30-38
18 Platelet alpha-granules are necessary for bone marrow-derived cell recruitment, tumor growth, and angiogenesis. Platelet accumulation Tumor Weight Blood Vessel Area
19 Platelets release SDF-1 and recruit circulating bone marrow-derived cells Stellos K, et al, Circulation, 2008 Massberg, Gawaz. J Exp Med 2006
20 In Vivo Recruitment of EPCs Through Plateletderived SDF-1 Mouse Ischemia Model (Microcirculation)
21 Platelets propagate differentiation of BMDC into mature endothelial cells Clot 4 days 7 days Langer et al., Circ Res, 2006
22 Platelets modulate function and differentiation of bone marrow-derived cells Angiogenesis Inflammation FASEB 2006; JEM 2006; Circ Res 2006; Circulation 2008; ATVB 2010; JMM 2010, Circ Res 2011
23 Anti-IgG CHO FcIgG2 kda CHO FcIgG2 GPVI kda GPVI Anti-GPVI CXCR4 CHO GPVI kda CHO GPVI Anti-SDF1 CHO SDF1 kda CHO SDF1 a Protein 1: GPVI-SDF1 c B Protein 2: 250 CXCR SDF-1 GPVI-SDF1 Linker b Human Fc GPVIreceptor Extracellular matrix (vascular injury) Ziegler et al., unpublished
24 Cell survival (% Live cells) Cell migration (migrated cells) Cell migration (migrated cells) a hsdf1 Ø 155nM GPVI-SDF1 155nM b Freshly Isolated cells Ø FcIgG2 TGFß1 hsdf1 PI 70% 12% 24% 15% 62% 48% 12% 28% 24% 23% 57% 43% 18% 28% 57% 28% 18% 24% Ziegler et al., unpublished Annexin V
25 c uncoated collagen I collagen + GPVI collagen + 1d 3d 5d 7d 10d 14d d vwf VEGFR-2 claudin 5 VE-cadherin Tie-2 PECAM-1 hcaec CD cells 14d Ziegler et al., unpublished
26 LV-systolic function (% FAC) infarct size (%I/AaR) a SDF1- GPVI 0d 2d 28d b c LAD ligation FcIgG2 0,1 0,4 0,7 0,1 0,4 0,7 diastole * Infarct size LV function FcIgG2 * 20 systole 10 Ziegler et al., unpublished FcIgG2 0 FcIgG2
27 capillaries/mm 2 stimulates angiogenese in myocardial infarction ctrl * Day SDF1- GPVI ctrl Ziegler et al., unpublished
28 stimulates angiogenese in myocardial infarction and preserves function CXCR4 Myocardial Repair Progenitor cells Bone marrow Heart
29 Thanks to Cardiology / Tübingen Peter Seizer Margitta Elvers Klaudia Posavec Tanja Schönberger Katharina Schuster Andreas May Konstantinos Stellos Oliver Borst Melanie Ziegler Pharmacology/ Ulm Th. Simmet Corimmun/ Martinsried Götz Münch Martin Ungerer Molecular Pathology / Tübingen Reinhard Kandolf Karin Klingel Physiology / Tübingen Florian Lang
30 Thank you for your attention! Tübingen, Germany
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