Our Contributions to Science

1. Defined the Cell Biology of von Willebrand Factor (VWF). Nothing was known about how VWF was made into the highly multimeric active adhesion molecules that are secreted. Our group’s first observation was that endothelial cells store VWF in granules called Weibel-Palade bodies. We discovered that VWF has a large prosequence that directs VWF multimerization and storage. In collaboration with colleagues from MIT, we produced VWF knockout mice that helped to elucidate VWF function in thrombosis and inflammation.

          Wagner DD, Olmsted JB, Marder VJ. Immunolocalization of von Willebrand protein in Weibel-Palade bodies of human endothelial cells. J Cell Biol. 1982 Oct;95(1):355-60. PubMed PMID: 6754744;
                  PMCID: PMC2112360.
          Wagner DD, Marder VJ. Biosynthesis of von Willebrand protein by human endothelial cells. Identification of a large precursor polypeptide chain. J Biol Chem. 1983 Feb 25;258(4):2065-7. PubMed                          PMID: 6600453.
          Sporn LA, Marder VJ, Wagner DD. Inducible secretion of large, biologically potent von Willebrand factor multimers. Cell. 1986 Jul 18;46(2):185-90. PubMed PMID: 3087627.
                   Denis C, Methia N, Frenette PS, Rayburn H, Ullman-Culleré M, Hynes RO, Wagner DD. A mouse model of severe von Willebrand disease: defects in hemostasis and thrombosis. Proc Natl Acad
                   Sci U.S A.    1998 Aug 4;95(16):9524-9. PubMed PMID: 9689113; PMCID: PMC21371. 

2. Uncovered the biological importance of P-selectin. P-selectin (then called PADGEM or GMP140) was known to be an activation antigen on platelets. Our group found that it is made also by endothelial cells and is stored with VWF in Weibel-Palade bodies. With our colleagues, we showed that it mediates adhesion of platelets to leukocytes. After preparing P-selectin knockout mice (at the time the first adhesion molecule knockout that lived), we found that P-selectin is crucial for leukocyte rolling in vivo. With our colleagues at MIT, we then made the first double knockout (P- and E-selectin) and defined the importance of the selectins in leukocyte recruitment to sites of inflammation and also to bone marrow.

        Bonfanti R, Furie BC, Furie B, Wagner DD. PADGEM (GMP140) is a component of Weibel-Palade bodies of human endothelial cells. Blood. 1989 Apr;73(5):1109-12. PubMed PMID: 2467701.
          Larsen E, Celi A, Gilbert GE, Furie BC, Erban JK, Bonfanti R, Wagner DD, Furie B. PADGEM protein: a receptor that mediates the interaction of activated platelets with neutrophils and monocytes.
                  Cell. 1989 Oct 20;59(2):305-12. PubMed PMID: 2478294.
          Mayadas TN, Johnson RC, Rayburn H, Hynes RO, Wagner DD. Leukocyte rolling and extravasation are severely compromised in P selectin-deficient mice. Cell. 1993 Aug 13;74(3):541-54. PubMed                      PMID: 7688665.
          Frenette PS, Mayadas TN, Rayburn H, Hynes RO, Wagner DD. Susceptibility to infection and altered hematopoiesis in mice deficient in both P- and E-selectins. Cell. 1996 Feb 23;84(4):563-74.                              PubMed PMID:8598043. 

3. Described new aspects of platelet biology. We observed that platelets behave in many aspects like leukocytes, and that they are actively involved in inflammatory responses. We found that platelets roll on the vessel wall in vivo and activated platelets expressing P-selectin promote acute and chronic inflammation. We observed that CD40L, a ligand important in immunity, enhances thrombosis. With our collaborators, we showed that platelets support angiogenesis and prevent hemorrhaging of blood vessels during inflammation.

          Frenette PS, Johnson RC, Hynes RO, Wagner DD. Platelets roll on stimulated endothelium in vivo: an interaction mediated by endothelial P-selectin. Proc Natl Acad Sci U S A. 1995 Aug                                        1;92(16):7450-4. PubMed PMID: 7543682; PMCID: PMC41357.
          André P, Prasad KS, Denis CV, He M, Papalia JM, Hynes RO, Phillips DR, Wagner DD. CD40L stabilizes arterial thrombi by a beta3 integrin--dependent mechanism. Nat Med. 2002 Mar;8(3):247-52.                      PubMed PMID: 11875495.
          Burger PC, Wagner DD. Platelet P-selectin facilitates atherosclerotic lesion development. Blood. 2003 Apr 1;101(7):2661-6. PubMed PMID: 12480714.
          Kisucka J, Butterfield CE, Duda DG, Eichenberger SC, Saffaripour S, Ware J, Ruggeri ZM, Jain RK, Folkman J, Wagner DD. Platelets and platelet adhesion support angiogenesis while preventing                          excessive hemorrhage. Proc Natl Acad Sci U S A. 2006 Jan 24;103(4):855-60. PubMed PMID: 16418262; PMCID: PMC1348013. 

4. Visualized thrombosis in vivo and determined specific functions of platelet adhesion molecules. We were the first laboratory to study blood vessels of knockout mice by intravital microscopy, which is now a widespread technique. This allowed us to undo several dogma, such as “fibrinogen is necessary for thrombus formation” and “VWF is important only under arterial shear.” Our work contributed to the understanding of the role of leukocyte-derived microparticles in coagulation and the importance of ADAMTS13 in reducing both thrombosis and inflammation.

          Ni H, Denis CV, Subbarao S, Degen JL, Sato TN, Hynes RO, Wagner DD. Persistence of platelet thrombus formation in arterioles of mice lacking both von Willebrand factor and fibrinogen. J Clin                            Invest.  2000 Aug;106(3):385-92. PubMed PMID: 10930441; PMCID: PMC314330.
          Hrachovinová I, Cambien B, Hafezi-Moghadam A, Kappelmayer J, Camphausen RT, Widom A, Xia L, Kazazian HH Jr, Schaub RG, McEver RP, Wagner DD. Interaction of P-selectin and PSGL-1                            generates microparticles that correct hemostasis in a mouse model of hemophilia A. Nat Med. 2003 Aug;9(8):1020-5. PubMed PMID: 12858167.
          Chauhan AK, Motto DG, Lamb CB, Bergmeier W, Dockal M, Plaimauer B, Scheiflinger F, Ginsburg D, Wagner DD. Systemic antithrombotic effects of ADAMTS13. J Exp Med. 2006 Mar 20;203(3):                          767- 76. PubMed PMID: 16533881; PMCID: PMC2118248.
          Brill A, Fuchs TA, Chauhan AK, Yang JJ, De Meyer SF, Köllnberger M, Wakefield TW, Lämmle B, Massberg S, Wagner DD. von Willebrand factor-mediated platelet adhesion is critical for deep vein                        thrombosis in mouse models. Blood. 2011 Jan 27;117(4):1400-7. PubMed PMID: 20959603; PMCID: PMC3056477. 


5. Discovered the importance of neutrophil extracellular traps (NETs) in thrombosis and sterile inflammation. Before our work, NETs formation was linked only to infection and autoimmune disease. We found that platelets bind to NETs and the enzyme PAD4, which decondenses chromatin to enable NET release, plays a profound role in DVT. In animal and human studies, we observed that cancer and diabetes promote NETosis, which may initiate thrombosis and negatively impacts wound healing.

          Fuchs TA, Brill A, Duerschmied D, Schatzberg D, Monestier M, Myers DD Jr, Wrobleski SK, Wakefield TW, Hartwig JH, Wagner DD. Extracellular DNA traps promote thrombosis. Proc Natl Acad Sci U                    S A. 2010 Sep 7;107(36):15880-5. PubMed PMID: 20798043; PMCID: PMC2936604.
          Demers M, Krause DS, Schatzberg D, Martinod K, Voorhees JR, Fuchs TA, Scadden DT, Wagner DD. Cancers predispose neutrophils to release extracellular DNA traps that contribute to cancer-                          associated thrombosis. Proc Natl Acad Sci U S A. 2012 Aug 7;109(32):13076-81. PubMed PMID: 22826226; PMCID: PMC3420209.
          Savchenko AS, Borissoff JI, Martinod K, De Meyer SF, Gallant M, Erpenbeck L, Brill A, Wang Y, Wagner DD. VWF-mediated leukocyte recruitment with chromatin decondensation by PAD4 increases                      myocardial ischemia/reperfusion injury in mice. Blood. 2014 Jan 2;123(1):141-8. PubMed PMID: 24200682; PMCID: PMC3879903.
          Wong SL, Demers M, Martinod K, Gallant M, Wang Y, Goldfine AB, Kahn CR, Wagner DD. Diabetes primes neutrophils to undergo NETosis, which impairs wound healing. Nat Med. 2015                                        Jul;21(7):815-9. PMID:26076037  PMCID: PMC4631120