title: Nucleotide-binding oligomerization domain 2 receptor is expressed in platelets and enhances platelet activation and thrombosis.
abstract: BACKGROUND
  Pattern recognition receptor nucleotide-binding oligomerization domain 2 (NOD2) is well investigated in immunity, but its expression and function in platelets has never been explored.
  
  
  RESULTS
  Using reverse transcription polymerase chain reaction and Western blot, we show that both human and mouse platelets express NOD2, and its agonist muramyl dipeptide induced NOD2 activation as evidenced by receptor dimerization. NOD2 activation potentiates platelet aggregation and secretion induced by low concentrations of thrombin or collagen, and clot retraction, as well. These potentiating effects of muramyl dipeptide were not seen in platelets from NOD2-deficient mice. Plasma from septic patients also potentiates platelet aggregation induced by thrombin or collagen NOD2 dependently. Using intravital microscopy, we found that muramyl dipeptide administration accelerated in vivo thrombosis in a FeCl3-injured mesenteric arteriole thrombosis mouse model. Platelet depletion and transfusion experiments confirmed that NOD2 from platelets contributes to the in vivo thrombosis in mice. NOD2 activation also accelerates platelet-dependent hemostasis. We further found that platelets express receptor-interacting protein 2, and provided evidence suggesting that mitogen activated-protein kinase and nitric oxide/soluble guanylyl cyclase/cGMP/protein kinase G pathways downstream of receptor-interacting protein mediate the role of NOD2 in platelets. Finally, muramyl dipeptide stimulates proinflammatory cytokine interleukin-1β maturation and accumulation in human and mouse platelets NOD2 dependently.
  
  
  CONCLUSIONS
  NOD2 is expressed in platelets and functions in platelet activation and arterial thrombosis, possibly during infection. To our knowledge, this is the first study on NOD-like receptors in platelets that link thrombotic events to inflammation.
authors.affiliation: From Key Laboratory of Molecular Medicine, Ministry of Education, and Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai, China (Si Zhang, Shenghui Zhang, L.H., L.Z., J.Y., L.C., Z.D.); Department of Internal Medicine, and Institute of Liver Disease, Fudan University Zhongshan Hospital, Shanghai, China (R.X.); Thoracic Oncology Research Laboratory, University of Pennsylvania, Philadelphia, (G.C.); Department of Internal Medicine, University of Kansas School of Medicine, Wichita (J.S.M.); and Department of Physiology and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA (S.P.K.).
authors.forename: Si
authors.initials: S
authors.lastname: Zhang
authors.affiliation: From Key Laboratory of Molecular Medicine, Ministry of Education, and Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai, China (Si Zhang, Shenghui Zhang, L.H., L.Z., J.Y., L.C., Z.D.); Department of Internal Medicine, and Institute of Liver Disease, Fudan University Zhongshan Hospital, Shanghai, China (R.X.); Thoracic Oncology Research Laboratory, University of Pennsylvania, Philadelphia, (G.C.); Department of Internal Medicine, University of Kansas School of Medicine, Wichita (J.S.M.); and Department of Physiology and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA (S.P.K.).
authors.forename: Shenghui
authors.initials: S
authors.lastname: Zhang
authors.affiliation: From Key Laboratory of Molecular Medicine, Ministry of Education, and Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai, China (Si Zhang, Shenghui Zhang, L.H., L.Z., J.Y., L.C., Z.D.); Department of Internal Medicine, and Institute of Liver Disease, Fudan University Zhongshan Hospital, Shanghai, China (R.X.); Thoracic Oncology Research Laboratory, University of Pennsylvania, Philadelphia, (G.C.); Department of Internal Medicine, University of Kansas School of Medicine, Wichita (J.S.M.); and Department of Physiology and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA (S.P.K.).
authors.forename: Liang
authors.initials: L
authors.lastname: Hu
authors.affiliation: From Key Laboratory of Molecular Medicine, Ministry of Education, and Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai, China (Si Zhang, Shenghui Zhang, L.H., L.Z., J.Y., L.C., Z.D.); Department of Internal Medicine, and Institute of Liver Disease, Fudan University Zhongshan Hospital, Shanghai, China (R.X.); Thoracic Oncology Research Laboratory, University of Pennsylvania, Philadelphia, (G.C.); Department of Internal Medicine, University of Kansas School of Medicine, Wichita (J.S.M.); and Department of Physiology and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA (S.P.K.).
authors.forename: Lili
authors.initials: L
authors.lastname: Zhai
authors.affiliation: From Key Laboratory of Molecular Medicine, Ministry of Education, and Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai, China (Si Zhang, Shenghui Zhang, L.H., L.Z., J.Y., L.C., Z.D.); Department of Internal Medicine, and Institute of Liver Disease, Fudan University Zhongshan Hospital, Shanghai, China (R.X.); Thoracic Oncology Research Laboratory, University of Pennsylvania, Philadelphia, (G.C.); Department of Internal Medicine, University of Kansas School of Medicine, Wichita (J.S.M.); and Department of Physiology and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA (S.P.K.).
authors.forename: Ruyi
authors.initials: R
authors.lastname: Xue
authors.affiliation: From Key Laboratory of Molecular Medicine, Ministry of Education, and Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai, China (Si Zhang, Shenghui Zhang, L.H., L.Z., J.Y., L.C., Z.D.); Department of Internal Medicine, and Institute of Liver Disease, Fudan University Zhongshan Hospital, Shanghai, China (R.X.); Thoracic Oncology Research Laboratory, University of Pennsylvania, Philadelphia, (G.C.); Department of Internal Medicine, University of Kansas School of Medicine, Wichita (J.S.M.); and Department of Physiology and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA (S.P.K.).
authors.forename: Jianqin
authors.initials: J
authors.lastname: Ye
authors.affiliation: From Key Laboratory of Molecular Medicine, Ministry of Education, and Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai, China (Si Zhang, Shenghui Zhang, L.H., L.Z., J.Y., L.C., Z.D.); Department of Internal Medicine, and Institute of Liver Disease, Fudan University Zhongshan Hospital, Shanghai, China (R.X.); Thoracic Oncology Research Laboratory, University of Pennsylvania, Philadelphia, (G.C.); Department of Internal Medicine, University of Kansas School of Medicine, Wichita (J.S.M.); and Department of Physiology and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA (S.P.K.).
authors.forename: Leilei
authors.initials: L
authors.lastname: Chen
authors.affiliation: From Key Laboratory of Molecular Medicine, Ministry of Education, and Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai, China (Si Zhang, Shenghui Zhang, L.H., L.Z., J.Y., L.C., Z.D.); Department of Internal Medicine, and Institute of Liver Disease, Fudan University Zhongshan Hospital, Shanghai, China (R.X.); Thoracic Oncology Research Laboratory, University of Pennsylvania, Philadelphia, (G.C.); Department of Internal Medicine, University of Kansas School of Medicine, Wichita (J.S.M.); and Department of Physiology and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA (S.P.K.).
authors.forename: Guanjun
authors.initials: G
authors.lastname: Cheng
authors.affiliation: From Key Laboratory of Molecular Medicine, Ministry of Education, and Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai, China (Si Zhang, Shenghui Zhang, L.H., L.Z., J.Y., L.C., Z.D.); Department of Internal Medicine, and Institute of Liver Disease, Fudan University Zhongshan Hospital, Shanghai, China (R.X.); Thoracic Oncology Research Laboratory, University of Pennsylvania, Philadelphia, (G.C.); Department of Internal Medicine, University of Kansas School of Medicine, Wichita (J.S.M.); and Department of Physiology and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA (S.P.K.).
authors.forename: Jozef
authors.initials: J
authors.lastname: Mruk
authors.affiliation: From Key Laboratory of Molecular Medicine, Ministry of Education, and Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai, China (Si Zhang, Shenghui Zhang, L.H., L.Z., J.Y., L.C., Z.D.); Department of Internal Medicine, and Institute of Liver Disease, Fudan University Zhongshan Hospital, Shanghai, China (R.X.); Thoracic Oncology Research Laboratory, University of Pennsylvania, Philadelphia, (G.C.); Department of Internal Medicine, University of Kansas School of Medicine, Wichita (J.S.M.); and Department of Physiology and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA (S.P.K.).
authors.forename: Satya P
authors.initials: SP
authors.lastname: Kunapuli
authors.affiliation: From Key Laboratory of Molecular Medicine, Ministry of Education, and Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai, China (Si Zhang, Shenghui Zhang, L.H., L.Z., J.Y., L.C., Z.D.); Department of Internal Medicine, and Institute of Liver Disease, Fudan University Zhongshan Hospital, Shanghai, China (R.X.); Thoracic Oncology Research Laboratory, University of Pennsylvania, Philadelphia, (G.C.); Department of Internal Medicine, University of Kansas School of Medicine, Wichita (J.S.M.); and Department of Physiology and Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA (S.P.K.). dingzr@fudan.edu.cn.
authors.forename: Zhongren
authors.initials: Z
authors.lastname: Ding
mesh_terms: D000119:Acetylmuramyl-Alanyl-Isoglutamine / Q000494:pharmacology; D000818:Animals; D016470:Bacteremia / Q000097:blood; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D003018:Clot Retraction / Q000502:physiology; D006152:Cyclic GMP / Q000097:blood; D019281:Dimerization; D006487:Hemostasis / Q000502:physiology; D006801:Humans; D007249:Inflammation / Q000097:blood*; D053583:Interleukin-1beta / Q000097:blood; D020935:MAP Kinase Signaling System / Q000502:physiology; D051379:Mice; D016688:Mice, Inbred NOD; D009569:Nitric Oxide / Q000097:blood; D053473:Nod2 Signaling Adaptor Protein / Q000819:agonists / Q000096:biosynthesis / Q000097:blood / Q000502:physiology*; D015539:Platelet Activation / Q000187:drug effects / Q000502:physiology*; D053475:Receptor-Interacting Protein Serine-Threonine Kinase 2 / Q000096:biosynthesis; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000096:biosynthesis; D015398:Signal Transduction / Q000502:physiology; D013927:Thrombosis / Q000097:blood*
chemical_list: C587672:IL1B protein, mouse; D053583:Interleukin-1beta; C429018:NOD2 protein, human; D053473:Nod2 Signaling Adaptor Protein; C495945:Nod2 protein, mouse; D009569:Nitric Oxide; D000119:Acetylmuramyl-Alanyl-Isoglutamine; C506253:RIPK2 protein, human; D053475:Receptor-Interacting Protein Serine-Threonine Kinase 2; D053422:Receptor-Interacting Protein Serine-Threonine Kinases; C506255:Ripk2 protein, mouse; D006152:Cyclic GMP
keywords: blood platelets; mitogen-activated protein kinases; nucleotide-binding oligomerization domain 2; receptor-interacting protein serine-threonine kinase 2; thrombosis
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