Platelet Neutrophil crosstalk in thrombotic disorders Dimitrios A. Stakos

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1 Platelet Neutrophil crosstalk in thrombotic disorders Dimitrios A. Stakos Assistant professor of Cardiology Cardiology Dept. & Laboratory of Molecular Hematology DUTH, Alexandroupolis

2 Aspects of the common ancestral functions between immunity and coagulation Massberg S. Nat Rev Immunol, 2013

3 Manual aspiration form IRA during primary PCI Αρχείο, Στάκος

4 PLATELETS AND INFLAMMATION (Neutrophil PLT aggregates) In patients with ACS Circulation, 1996

5 PLATELETS AND INFLAMMATION (Neutrophil PLT aggregates) In patients with ACS Circulation, 1996

6 high mobility group box 1

7 Brinkmann V, et al. Neutrophil extracellular traps kill bacteria. Science 2004 Neutrophil derived structures composed of extruded DNA, decorated with antimicrobial proteins

8 NET s function High local concentration of antimicrobial components Kill microbes extracellularly independent of phagocytic uptake Physical barrier that prevents further spread of the pathogens Delivery of granule proteins into NETs (keep injurious proteins proteases from diffusing)

9 Brinkmann V. J Cell Biol Firm attachment to the substratum Disintegration of membranes mixture Cell contraction membrane rupture

10 TF exposure on NETs in Sepsis.evidence for the implication of this process in coagulopathy and inflammatory response in sepsis Kambas K, et al. PLoS One Kambas K. et al. Front Immunol. 2012

11 TF exposure on NETs in septic patients.evidence for the implication of this process in coagulopathy and inflammatory response in sepsis Kourtzelis I, et al. Blood Kambas K, et al. PLoS One. 2012

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14 ctr plasma Ενδοκυττάριος TF IRA Ενδοκυττάριος TF nira

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16 Activated platelets Ενδοκυττάριος TF + + TF bearing NETs IRA plasma

17 Stakos D. et al. Eur Heart J. 2015

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19 Αγγειακό τοίχωμα Αιμοπετάλια PMN

20 Ενδοκυττάριος TF ΑυτοφαγίαTF/έναρξη NETosis PMN PLT αλληλεπίδραση

21 TF thrombin axis: σταθεροποίηση θρόμβου Έκφραση TF πάνω σε ΝΕ

22 GPVI PLT Αρχικός θρόμβος GPIb vwf collagen Ρήξη κάψας PLT Ρήξη πλάκας/παθολογική θρόμβωση Αιμόσταση

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24 Fibrin scattered inflammatory cells

25 Co-incubation of stents with control neutrophils DES BMS TF IL17 IL17 NE NE NE BMS, DES, BVS

26 Stent-induced NETs in fibroblast cultures Control α-sma (+)

27 Co-incubation of stents with IRA-plasma activated neutrophils

28 Platelet mediators release NETs Vacant cells release NETs

29 high mobility group box 1

30 Platelets induce NET formation Activated platelets induce NET formation in acute lung injury (ALI) Thrombin Receptor Activating Peptide (PAR 1 agonist) Caudrillier A, et al. J Clin Invest. 2012

31 Stakos D, et al. Eur Heart J * Thrombin DNase * NET scaffold integrity is imperative for TF functionality 3 D conformation / TFPI degradation (NE)

32 Semeraro F, et al. Blood Clark SR, et al. Nat Med (sepsis)

33 Clot stability under sheer stress Pure fibrin Fibrin containing 50g/mlDNA Fibrin containing 100 g/ml DNA Pa: pascals Clot stability under sheer stress in the presence of 300 g/ml histone and Clot stability under sheer stress in the presence 300 g/ml histone g/ml DNA. J. Biol. Chem. 2013

34 sections NETs in Chronic thromboelbolic pulmonary hypertension Tissue characterization

35 NETs in Chronic thromboelbolic pulmonary hypertension Endothelin 1

36 Modulators / Inhibitors

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38 mir binds following Watson Crick base pairing seed region 3 UTR mir guides RISC to target mrna RISC Mature mrna coding sequence 3 UTR poly A tail Cap 5 UTR Translation repression

39 IL / FeCl arterial thrombosis models PMNs expressing TF is the main source of TF needed for platelet thrombus formation and fibrin generation following arterial injury in vivo. Darbousset R, et al. Blood 2012

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41 Pathogen elimination PLT Tissue Factor Thrombosis Sepsis VEGF IL 1β Angiogenesis Plaque vulnerability Cancer biology Disease specific NET markers NETomics as diagnostic tools EPITOPES F. Mediterranean fever Gout Colitis Matrix metalloproteinases (MMPs) IL 17 Plaque instability/rupture Plaque stabilization Fibrosis Wound healing Tissue remodeling Pulmonary fibrosis Inflammatory Bowel disease Restenosis after stenting

42 Internal medicine, Hematology Σταύρος Γιαγλής, Biology Σταύρος Ραφαήλ, Mol Biology Ιωάννης Κουρτζέλης, Biology Αποστολία Τζέκου, Mol. Biology Ιωάννης Μητρούλης, Internal Medicine Στέλλα Αρελάκη, Pathologist Immunology Mol Biol, BSc. MRes, Mol Biol, Postdoc, Cardiology Technician Biopathology Internal medicine Immunology Mol Biol, Master Mol Biol, Master Mol Biol, PhD