Purinergic Thromboregulation

嘌呤能血栓调节

• 批准号: 7851207
• 负责人: SIMON C. ROBSON
• 金额: $ 63.75万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7851207
• 关键词: ADP Receptors; ATP Receptors; Accounting; Adenosine; Adoptive Transfer; Agonist; Anti-Inflammatory Agents; Anti-inflammatory; Apolipoprotein A-II; Binding; Blood Cells; Blood Circulation; Blood Platelets; Blood Vessels; Bone Marrow Transplantation; Cations; Cell membrane; Cells; Ceramides; Chimeric Proteins; Cholesterol; Coagulation Process; Coupled; Data; Deposition; Endothelial Cells; Endothelium; Enzymes; Family; Fibrin; GTP-Binding Proteins; Generations; Growth Factor Receptors; Hemostatic function; High Density Lipoproteins; Human; Hydrolysis; Immune; Individual; Injury; Integrins; Kinetics; Knockout Mice; Leukocytes; Life; Link; Lipids; Lipoprotein (a); Lipoproteins; Lymphocyte; Macromolecular Complexes; Mediating; Mediator of activation protein; Membrane Microdomains; Membrane Proteins; Monitor; Mus; Mutant Strains Mice; NTPDase2; Nucleosides; Nucleotides; P-Selectin; P2X-receptor; Pericytes; Phase; Plasma; Platelet Activation; Platelet aggregation; Play; Post-Translational Protein Processing; Process; Property; RanBMP-90; Receptor Signaling; Regulatory T-Lymphocyte; Relative (related person); Role; Scaffolding Protein; Signal Transduction; Site; Smooth Muscle Myocytes; Sphingomyelins; Testing; Thromboplastin; Thrombosis; Thrombus; Transgenic Mice; Transgenic Organisms; Vascular Endothelium; Video Microscopy; Wild Type Mouse; Work; acid sphingomyelinase; biochemical evolution; ectoADPase; ectoATPase; extracellular; immune function; in vivo; inhibitor/antagonist; insight; killer T cell; mutant; nucleoside triphosphate; palmitoylation; receptor; research study; response; vascular inflammation

Extracellular nucleotides (ATP, ADP and UTP) are known to activate type-2 purinergic (P2Y and P2X) receptors on platelets, endothelium and immune cells to induce vascular inflammation and thrombosis. Cellular ectonucleotidases hydrolyze P2-mediators to down regulate P2 receptor-signaling responses. CD39 is the dominant vascular, T regulatory and Natural Killer (NK)T cell ectonucleotidase/NTPDase and plays a crucial role in the ectonucleotidase cascade, involving CD73, to generate adenosine. This nucleoside has important antithrombotic and anti-inflammatory properties. The roles of CD39L1 (a preferential ecto-ATPase), expressed by adventitial cells, and CD39L3 (functionally akin to CD39) expressed by vascular endothelium, on thrombus formation are completely unexplored in vivo. Specific Aim 1: Study functional impacts of the three vascular CD39/NTPDases on thrombus formation in vivo. a) Evaluate and compare accumulation of CD39, CD39L1 and CD39L3 in generated thrombi, with platelet sequestration, associated tissue factor and fibrin deposition. b) Contrast the impact of vascular Cd39L1 and/or Cd39L3 deletion vs. global absence of Cd39 on arteriolar thrombus formation and platelet activation in mutant mice. Specific Aim 2: Thromboregulatory functions of immune cells in vivo a) Perform adoptive transfer of T regulatory cells expressing Cd39 to determine the role of immune regulatory cells on platelet activation and thrombus formation in vivo. b) Test mice following adoptive transfer of isolated NKT cells expressing Cd39 and Cd73 to determine impact on thrombus formation in vivo. Specific Aim 3: Association of CD39 with membrane proteins and lipids. Palmitoylation at the N-terminus targets CD39 to lipid rafts within cell membranes that are linked to microparticle formation. The C-terminus of CD39 binds apo-lipoprotein A2 and incorporates CD39 into High Density Lipoproteins (HDL). a) Explore structural and functional interactions of CD39 within lipid rafts. b) Examine how palmitoylation and other post-translational modifications of CD39 impact association with cholesterol and sphingomyelin. c) Determine effects of activation of acid sphingomyelinase and ceramide generation on CD39 bioactivity in endothelial cells, lymphocytes and their derived microparticles. d) Study thrombus formation in hyperlipidemic Cd39 mutant mice. This work will provide insights into the central thromboregulatory mechanisms impacting platelet activation and vascular injury in vivo.

已知细胞外核苷酸（ATP、ADP 和 UTP）可激活血小板、内皮和免疫细胞上的 2 型嘌呤能（P2Y 和 P2X）受体，从而诱导血管炎症和血栓形成。细胞核酸外切酶水解 P2 介体以下调 P2 受体信号传导反应。 CD39 是主要的血管、T 调节和自然杀伤 (NK)T 细胞核酸外切酶/NTPDase，在涉及 CD73 的核酸外切酶级联中发挥着至关重要的作用，以生成腺苷。这种核苷具有重要的抗血栓和抗炎特性。外膜细胞表达的 CD39L1（一种优先外源 ATP 酶）和血管内皮细胞表达的 CD39L3（功能类似于 CD39）在血栓形成中的作用在体内完全未被探索。具体目标 1：研究三种血管 CD39/NTPDase 对体内血栓形成的功能影响。 a) 评估并比较生成的血栓中 CD39、CD39L1 和 CD39L3 的积累、血小板隔离、相关组织因子和纤维蛋白沉积。 b) 对比血管 Cd39L1 和/或 Cd39L3 缺失与 Cd39 整体缺失对突变小鼠小动脉血栓形成和血小板活化的影响。具体目标2：体内免疫细胞的血栓调节功能a)对表达Cd39的T调节细胞进行过继转移，以确定免疫调节细胞在体内对血小板活化和血栓形成的作用。 b) 在过继转移表达 Cd39 和 Cd73 的分离的 NKT 细胞后测试小鼠，以确定对体内血栓形成的影响。具体目标 3：CD39 与膜蛋白和脂质的关联。 N 末端的棕榈酰化将 CD39 靶向细胞膜内与微粒形成相关的脂筏。 CD39 的 C 末端结合载脂蛋白 A2 并将 CD39 整合到高密度脂蛋白 (HDL) 中。 a) 探索脂筏内 CD39 的结构和功能相互作用。 b) 检查 CD39 的棕榈酰化和其他翻译后修饰如何影响与胆固醇和鞘磷脂的关联。 c)确定酸性鞘磷脂酶的激活和神经酰胺的生成对内皮细胞、淋巴细胞及其衍生微粒中CD39生物活性的影响。 d) 研究高脂血症 Cd39 突变小鼠的血栓形成。这项工作将深入了解影响体内血小板活化和血管损伤的中央血栓调节机制。

项目成果

Adenosine: tipping the balance towards hepatic steatosis and fibrosis.

腺苷：使肝脏脂肪变性和纤维化的平衡倾斜。

• DOI: 10.1016/j.jhep.2010.02.009
• 发表时间: 2010
• 期刊: Journal of hepatology
• 影响因子: 25.7
• 作者: Robson,SimonC;Schuppan,Detlef
• 通讯作者: Schuppan,Detlef

High-dimensional analysis of the adenosine pathway in high-grade serous ovarian cancer.

• DOI: 10.1136/jitc-2020-001965
• 发表时间: 2021-03
• 期刊: Journal for immunotherapy of cancer
• 影响因子: 10.9
• 作者: Bareche Y;Pommey S;Carneiro M;Buisseret L;Cousineau I;Thebault P;Chrobak P;Communal L;Allard D;Robson SC;Mes-Masson AM;Provencher D;Lapointe R;Stagg J
• 通讯作者: Stagg J

CD73 is a phenotypic marker of effector memory Th17 cells in inflammatory bowel disease.

• DOI: 10.1002/eji.201242623
• 发表时间: 2012-11
• 期刊: EUROPEAN JOURNAL OF IMMUNOLOGY
• 影响因子: 5.4
• 作者: Doherty, Glen A.;Bai, Aiping;Hanidziar, Dusan;Longhi, Maria S.;Lawlor, Garrett O.;Putheti, Prabhakar;Csizmadia, Eva;Nowak, Martina;Cheifetz, Adam S.;Moss, Alan C.;Robson, Simon C.
• 通讯作者: Robson, Simon C.

CD39 limits P2X7 receptor inflammatory signaling and attenuates sepsis-induced liver injury.

• DOI: 10.1016/j.jhep.2017.05.021
• 发表时间: 2017-10
• 期刊: Journal of hepatology
• 影响因子: 25.7
• 作者: Savio LEB;de Andrade Mello P;Figliuolo VR;de Avelar Almeida TF;Santana PT;Oliveira SDS;Silva CLM;Feldbrügge L;Csizmadia E;Minshall RD;Longhi MS;Wu Y;Robson SC;Coutinho-Silva R
• 通讯作者: Coutinho-Silva R

NTPDase1 governs P2X7-dependent functions in murine macrophages.

• DOI: 10.1002/eji.200939741
• 发表时间: 2010-05
• 期刊: EUROPEAN JOURNAL OF IMMUNOLOGY
• 影响因子: 5.4
• 作者: Levesque, Sebastien A.;Kukulski, Filip;Enjyoji, Keiichi;Robson, Simon C.;Sevigny, Jean
• 通讯作者: Sevigny, Jean