Antiphospholipid antibodies and lupus: new molecular targets for treatment

抗磷脂抗体和狼疮：治疗的新分子靶点

• 批准号: 7889648
• 负责人: SILVIA S PIERANGELI
• 金额: $ 34.43万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7889648
• 关键词: ANXA2 gene; Affect; Affinity; Alteplase; Animal Model; Antibodies; Anticoagulation; Antigens; Antiphospholipid Antibodies; Antiphospholipid Syndrome; Aorta; Binding; Blood Platelets; Carotid Arteries; Cell Communication; Cell membrane; Cells; Clinical; Coagulants; Complex; Disease susceptibility; Endothelial Cells; Event; Family; IL6 gene; IL8 gene; Immunodominant Epitopes; Immunosuppression; In Vitro; Individual; Inflammatory; Laser Scanning Microscopy; Lead; Ligands; Lipopolysaccharides; Low Density Lipoprotein Receptor; Lupus; Mediating; Modality; Molecular Target; Mononuclear; Morbidity - disease rate; Mus; Myelogenous; Names; Nature; Patients; Peritoneal; Phenotype; Phosphorylation; Plasminogen; Platelet aggregation; Pregnancy loss; Prevention; Proteins; Quantum Dots; Signal Transduction; System; Systemic Lupus Erythematosus; Therapeutic Agents; Thromboplastin; Thrombosis; Thrombus; Up-Regulation; Vascular Cell Adhesion Molecule-1; Vascular Endothelial Growth Factors; apolipoprotein E receptor 2; cell type; crosslink; cytokine; human MAPK14 protein; in vivo; intercellular cell adhesion molecule; member; monocyte; mortality; mutant; nanocrystal; prevent; public health relevance; receptor; toll-like receptor 4; two-photon

DESCRIPTION (provided by applicant): Antiphospholipid (aPL) antibodies (Abs) are associated with thrombosis and pregnancy loss in patients with systemic lupus erythematosus (SLE) and antiphospholipid syndrome (APS). Thrombosis is an important cause of morbidity and mortality in APS and SLE patients with aPL Abs. APL Abs antibodies recognize domain I (DI) of 22glycoprotein I (22GPI). 22GPI binds to target cells [i.e.: endothelial cells (EC), platelets, monocytes] through domain V and trigger an intracellular signaling and a pro-coagulant and pro-inflammatory phenotype [i e.: expression of tissue factor (TF), intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), upregulation of cytokines [IL1b, IL6, IL8, TNF-a, vascular endothelial growth factor (VEGF)]. There is strong evidence that annexin A2, a receptor for tissue plasminogen activator (tPA) and plasminogen, and toll-like receptor 4 (TLR-4), a receptor for bacterial lipopolysaccharide (LPS) and apolipoprotein E receptor 2' (apoER2') may bind 22GPI and trigger intracellular signaling in target cells. Hence, the receptor(s) for 22GPI in target cells may involve more than one protein that would ultimately cluster or cross-link with aPL/a22GPI Abs and initiate intracellular signaling events, leading to a pro-thrombotic diathesis. We hypothesize that aPL/anti-22GPI pathogenic effects may be abrogated by inhibiting the specific binding of aPL/a22GPI Abs to DI of 22GPI or by blocking the interaction of 22GPI with the receptor(s) proteins recognized by 22GPI on target cells. We will examine this question utilizing various in vitro and in vivo approaches. We will first examine whether a TLR-4 ligand antagonist, anti-TLR-4 antibodies, anti-annexin A2 Abs or soluble binding domain 1 (BD1) of apoER2', or a common antagonist to members of the LDL receptor family named receptor associated protein (RAP) affect aPL-mediated upregulation of TF, ICAM-1, cytokines and p38 mitogen activated protein kinase (p38 MAPK) phosphorylation in EC and activation of monocytes and platelets. Then, we will examine the effects aPL/a22GPI Abs on thrombus formation, VCAM-1 and TF expression in aortas of mice (using quantum dot nano crystals and two-photon excitation laser scanning microscopy), cytokine upregulation (using a Multiplex/Luminex platform system), TF function in carotid artery homogenates and mononuclear peritoneal cells and platelet aggregation, in annexin A2, in myeloid differentiation factor (MyD)88 - an intracellular protein downstream from TLR-4, in apoER2' deficient mice and in normal mice treated with the specific abs/antagonists and aPL/a22GPI antibodies. In addition, we will study the ability of pegylated wild-type DI of 22GPI and some of its mutants - that have been shown to bind aPL/a22GPI with various affinities and inhibit some aPL-mediated effects- to affect the pathogenic effects of aPL/a22GPI Abs in vitro in various target cells and in mice. These studies will provide significant information on the nature of the interactions of 22GPI /aPL/a22GPI complexes with target cells in vitro and in vivo and will help to devise new targeted modalities for treatment/prevention of thrombosis in SLE patients with aPL/a22GPI Abs. PUBLIC HEALTH RELEVANCE: Antiphospholipid (aPL) antibodies (Abs) are associated with thrombosis and pregnancy loss in patients with lupus and the antiphospholipid syndrome (APS) and those clinical manifestations are an important cause of morbidity and mortality in individuals affected. Here we propose to target the interactions of aPL Abs with their antigen on various types of cells as a means to ameliorate aPL Abs-pathogenic effects in vitro and in animal models. These studies may help to devise new more specific and less harmful modalities than the anticoagulation or general immunosuppression currently used to treat and prevent thrombosis in patients with aPL Abs.

描述（由申请人提供）：抗磷脂（APL）抗体（ABS）与全身性红斑狼疮（SLE）和抗磷脂综合征（APS）患者的血栓形成和妊娠损失有关。血栓形成是APS和APL ABS的APS和SLE患者发病率和死亡率的重要原因。 APL ABS抗体识别22glycopoyin I（22GPI）的结构域I（DI）。 22GPI通过结构域V与靶细胞（即内皮细胞（EC），血小板，单核细胞]结合，并触发细胞内信号传导和促凝和促凝的和促启动的表型[I。e。e。e。：：细胞因子[IL1B，IL6，IL8，TNF-A，血管内皮生长因子（VEGF）]。 There is strong evidence that annexin A2, a receptor for tissue plasminogen activator (tPA) and plasminogen, and toll-like receptor 4 (TLR-4), a receptor for bacterial lipopolysaccharide (LPS) and apolipoprotein E receptor 2' (apoER2') may bind 22GPI and trigger intracellular signaling in target cells.因此，目标细胞中22GPI的受体可能涉及多种蛋白质，最终会与APL/A22GPI ABS聚类或交联并启动细胞内信号传导事件，从而导致促血栓形成透明的素质。我们假设APL/抗22GPI致病作用可以通过抑制APL/A22GPI ABS与22GPI的DI的特异性结合或通过阻断22GPI与22GPI识别的22GPI识别靶细胞对受体蛋白的相互作用的特异性结合。我们将利用各种体外和体内方法研究这个问题。 We will first examine whether a TLR-4 ligand antagonist, anti-TLR-4 antibodies, anti-annexin A2 Abs or soluble binding domain 1 (BD1) of apoER2', or a common antagonist to members of the LDL receptor family named receptor associated protein (RAP) affect aPL-mediated upregulation of TF, ICAM-1, cytokines and p38 mitogen activated protein EC中的激酶（p38 MAPK）磷酸化以及单核细胞和血小板的激活。 Then, we will examine the effects aPL/a22GPI Abs on thrombus formation, VCAM-1 and TF expression in aortas of mice (using quantum dot nano crystals and two-photon excitation laser scanning microscopy), cytokine upregulation (using a Multiplex/Luminex platform system), TF function in carotid artery homogenates and mononuclear peritoneal cells and platelet aggregation,在Annexin A2中，在髓样分化因子（MyD）88中 - TLR -4下游的细胞内蛋白，在ApoER2'缺陷小鼠中以及用特定ABS/拮抗剂和A22GPI抗体治疗的正常小鼠中。此外，我们将研究22GPI及其某些突变体的卵巢式野生型DI的能力 - 这些突变体已证明可以结合具有不同亲和力的APL/A22GPI，并抑制某些APL介导的效应 - 影响A22GPI ABS在各种靶细胞中的APP/A22GPI ABS的致病作用。这些研究将提供有关22GPI/APL/A22GPI复合物与靶细胞在体外和体内相互作用的性质的重要信息，并将有助于设计新的靶向方式，以治疗/预防A22GPI ABS的SLE患者的血栓形成。 公共卫生相关性：狼疮和抗磷脂综合征（AP）患者的抗磷脂（APL）抗体（ABS）与血栓形成和妊娠丧失有关，而临床表现是受影响个体的发病率和死亡率的重要原因。在这里，我们建议将APL ABS与它们的抗原相互作用在各种类型的细胞上的相互作用，以此来改善体外和动物模型中的APL ABS-PARNOGONIC效应。这些研究可能有助于与目前用于治疗和预防APL ABS患者血栓形成的抗凝抗凝或一般免疫抑制相比，将新的更特异性和危害方式更为有害。