Estradiol-Induced Sialyation Of NCAM Prevents Leucocyte:Endothelial Adhesion

雌二醇诱导的 NCAM 唾液酸化防止白细胞：内皮粘附

• 批准号: 7827373
• 负责人: FREDERICK NAFTOLIN
• 金额: $ 49.65万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7827373
• 关键词: Address; Adhesions; Affect; Area; Arterial Fatty Streak; Arteries; Atherosclerosis; Binding; Biological Assay; Blood Platelets; Blood Vessels; CCL2 gene; Cadherins; Cardiovascular system; Carotid Artery Ulcerating Plaque; Cause of Death; Cell Communication; Cell Line; Cells; Clinical; Clinical Research; Development; Documentation; Endothelial Cells; Endothelium; Enzymes; Equilibrium; Erythrocytes; Estradiol; Estrogen Receptors; Estrogens; Extracellular Domain; Foam Cells; Fulvestrant; Glycocalyx; Goals; Health; Healthcare; Heart; Heart Diseases; Hematological Disease; Hormonal; Human; Individual; Inflammation; Inflammatory; Lead; Leukocytes; Life; Lung; Lymphatic; Mediating; Menopause; Methods; Modeling; Myocardial Infarction; NCAM1 gene; Neural Cell Adhesion Molecules; New Agents; Organ; Ovarian; Parents; Pathogenesis; Pathway interactions; Personal Expenditure; Primates; Process; Production; Protein Isoforms; Rattus; Reaction; Regulation; Research Design; Reverse Transcriptase Polymerase Chain Reaction; Risk; Role; ST8Sia II; Sialic Acids; Sialyltransferases; Signal Transduction; Small Interfering RNA; Surface Antigens; Testing; Time; Umbilical vein; Valproic Acid; Vascular Endothelial Cell; Vascular Endothelium; Veins; Western Blotting; Woman; analog; atherogenesis; base; body system; brain tissue; chemokine; clinical practice; design; immunocytochemistry; improved; macrophage; monocyte; novel; prevent; protective effect; public health relevance; sialylation; vascular inflammation

DESCRIPTION (provided by applicant): This proposal addresses broad Challenge Area (04) Clinical Research and specific Challenge topic, 04- HL-103: Assess the role of leukocyte interaction with platelets, erythrocytes, and endothelium in the pathogenesis of heart, lung, and blood diseases. The cadherin Neural Cell Adhesion Molecule (NCAM) is key in leukocyte adhesion to endothelial cells. Upon signaling by chemokines such as MCP1 leukocytes are drawn toward endothelial cells. Appositional interactions ("zippering") between NCAM molecules from the leukocyte and endothelial cell forms a tether between them and allows the leukocyte's cell surface antigens to interact with those of the endothelial cell. This is followed by cell:cell interactions such as diapedesis. The resulting sub-endothelial inflammatory processes can result in atheroma formation and atherosclerosis. However, this sequence may be blocked by the insertion of the NCAM by sialic acid decorated isoforms termed polysialated NCAM (PSA-NCAM). These have poly hydrated ¿2,8 sialic acid chains in the extracellular domain that interfere with the zipper adhesion to NCAM. We have documented the presence of PSA-NCAM in the glycocalyx of human vascular endothelial cells in arteries, veins and lymphatics of all organs (10 organ systems) studied. We also have shown that expression of the two required polysialyltransferases (ST8Sia II/STX and ST8Sia IV/PST) is estrogen receptor-mediated. Thus, the enzymatic replacement of NCAM with PSA-NCAM depletes the parent NCAM molecules, so that the adhesion (NCAM):non- adhesion (PSA-NCAM) balance is highly leveraged. In the case of the vascular endothelium, the result may be minimization of leukocyte adhesion and sub endothelial inflammation, which would diminish the formation of atheromas and atherosclerosis. In previous studies we have shown that estradiol induces PST and STX in rat brain tissue and increases PSA- NCAM at the expense of NCAM in human umbilical vein endothelial cells (HUVEC). The estrogen receptor blocker fulvestrant had the opposite effect. In this proposal, using cardiovascular-relevant cells [human leukocytes (THP1) and human arterial endothelial cells (HCAEC)] we will examine estrogen-regulated leukocyte adhesion: We will determine the NCAM:PSA-NCAM ratio induced by estradiol and fulvestrant, their effects on sialylation enzymes and their effect on leukocyte-endothelial adhesion. We will compare the results with effects of the Ras pathway inducer valproic acid which is known to induce PSA-NCAM. Further, the effects of siRNA inhibition of STX and PST on the NCAM:PSA-NCAM balance and on leukocyte adhesion to endothelial cells will also be investigated. In this manner, we will test whether estrogen's cardioprotective effects could be due to increased NCAM sialylation, leading to decreased leukocyte adhesion and sub-endothelial inflammation. Impact: Heart disease is the single greatest cause of death and results in staggering healthcare expenditures and personal suffering. Finding new ways to protect against atherosclerosis is imperative for the health of the nation. Because of its importance to atherogenesis, understanding of the regulation of leukocyte-endothelial cell interactions is of the greatest clinical importance. This proposal will test the role of estrogen in preventing leukocyte adhesion that is a premonitory step toward atheroma formation. Such action would be consistent with the well- documented cardio-protective effects of estrogen in experimental and clinical studies. If this proposal is successful in exposing the mechanism by which estrogen affects leukocyte adhesion to endothelial cells it may be possible to alter already available treatments to avoid atherosclerosis and develop novel preventative treatments specifically aimed at regulating NCAM sialylation in clinical cardioprotection. Arteriosclerotic heart disease, the number one killer in the USA, starts from blood vessel inflammation that ultimately puts the individual at risk of a heart attack. This proposal will explore whether estrogen protects against atherosclerosis through production of specific, estrogen-regulated enzymes that inhibit inflammatory cells from entering into blood vessels. This proposal may improve the application of anti-atherosclerosis agents and lead to development of new agents to protect blood vessels from atherosclerosis and its complications. PUBLIC HEALTH RELEVANCE: Arteriosclerotic heart disease, the number one killer in the USA, starts from blood vessel inflammation that ultimately puts the individual at risk of a heart attack. This proposal will explore whether estrogen protects against atherosclerosis through production of specific, estrogen-regulated enzymes that inhibit inflammatory cells from entering into blood vessels. This proposal may improve the application of anti-atherosclerosis agents and lead to development of new agents to protect blood vessels from atherosclerosis and its complications.

描述（由申请人提供）： 本提案涉及广泛的挑战领域（04）临床研究和特定的挑战主题04- HL-103：评估白细胞与血小板、红细胞和内皮细胞相互作用在心脏、肺和血液疾病发病机制中的作用。钙粘蛋白神经细胞粘附分子（NCAM）是白细胞粘附内皮细胞的关键。在通过趋化因子如MCP 1发出信号后，白细胞被吸引向内皮细胞。来自白细胞和内皮细胞的NCAM分子之间的并置相互作用（“拉链”）在它们之间形成系链，并允许白细胞的细胞表面抗原与内皮细胞的细胞表面抗原相互作用。随后是细胞与细胞的相互作用，如渗出。所产生的内皮下炎症过程可导致动脉粥样硬化形成和动脉粥样硬化。然而，该序列可通过被称为多唾液酸化NCAM（PSA-NCAM）的唾液酸修饰的同种型插入NCAM来阻断。这些在细胞外结构域中具有多水合的<$2，8唾液酸链，其干扰拉链与NCAM的粘附。我们已经记录了PSA-NCAM在所研究的所有器官（10个器官系统）的动脉、静脉和动脉中的人血管内皮细胞的糖萼中的存在。我们还表明，表达所需的两个多聚唾液酸转移酶（ST 8 Sia II/STX和ST 8 Sia IV/PST）是雌激素受体介导的。因此，用PSA-NCAM酶促取代NCAM耗尽了亲本NCAM分子，使得粘附（NCAM）：非粘附（PSA-NCAM）平衡被高度利用。在血管内皮的情况下，结果可能是白细胞粘附和内皮下炎症的最小化，这将减少动脉粥样硬化和动脉粥样硬化的形成。在以前的研究中，我们已经表明雌二醇诱导大鼠脑组织中的PST和STX，并增加PSA-NCAM，而人脐静脉内皮细胞（HUVEC）中的NCAM减少。雌激素受体阻滞剂氟维司群则有相反的作用。在本提案中，使用心血管相关细胞[人白细胞（THP 1）和人动脉内皮细胞（HCAEC）]，我们将检查雌激素调节的白细胞粘附：我们将确定雌二醇和氟维司群诱导的NCAM：PSA-NCAM比率，它们对唾液酸化酶的影响以及它们对白细胞-内皮细胞粘附的影响。我们将比较结果与已知可诱导PSA-NCAM的Ras途径诱导剂丙戊酸的作用。此外，还将研究STX和PST的siRNA抑制对NCAM：PSA-NCAM平衡和对白细胞粘附至内皮细胞的影响。以这种方式，我们将测试雌激素的心脏保护作用是否可能是由于增加NCAM唾液酸化，导致白细胞粘附和内皮下炎症减少。影响：心脏病是死亡的最大原因，导致惊人的医疗支出和个人痛苦。寻找预防动脉粥样硬化的新方法对国家的健康至关重要。由于其在动脉粥样硬化形成中的重要性，了解白细胞-内皮细胞相互作用的调节具有最重要的临床意义。这项建议将测试雌激素在防止白细胞粘附中的作用，白细胞粘附是动脉粥样硬化形成的前兆。这种作用与实验和临床研究中雌激素的心脏保护作用是一致的。如果这个提议成功地揭示了雌激素影响白细胞与内皮细胞粘附的机制，那么就有可能改变现有的治疗方法以避免动脉粥样硬化，并开发新的预防性治疗方法，特别是在临床心脏保护中调节NCAM唾液酸化。动脉粥样硬化性心脏病是美国的头号杀手，它始于血管炎症，最终使患者面临心脏病发作的风险。这项提案将探讨雌激素是否通过产生特定的雌激素调节的酶来防止动脉粥样硬化，这些酶抑制炎症细胞进入血管。这一建议可能会改善抗动脉粥样硬化药物的应用，并导致开发新的药物，以保护血管免受动脉粥样硬化及其并发症。 公共卫生关系：动脉粥样硬化性心脏病是美国的头号杀手，它始于血管炎症，最终使患者面临心脏病发作的风险。这项提案将探讨雌激素是否通过产生特定的雌激素调节的酶来防止动脉粥样硬化，这些酶抑制炎症细胞进入血管。这一建议可能会改善抗动脉粥样硬化药物的应用，并导致开发新的药物，以保护血管免受动脉粥样硬化及其并发症。