The role of neutrophil recruitment in arterial diseases

中性粒细胞募集在动脉疾病中的作用

• 批准号: 8153357
• 负责人: YUQING HUO
• 金额: $ 4.9万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-08 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8153357
• 关键词: Affect; Angioplasty; Antibodies; Apolipoprotein E; Area; Arterial Fatty Streak; Arterial Injury; Arteries; Atherosclerosis; Binding; Blood Platelets; Breeding; Cell Adhesion Molecules; Cells; Collagen; Development; Diet; Disease; Disease model; Endothelium; Enzymes; Evans blue stain; Genes; Glycoproteins; Goals; Golgi Apparatus; Homeostasis; Homing; Inflammatory; Injury; Knock-out; L-Selectin; Lead; Lesion; Leukocytes; Ligands; Link; Lipids; Lymphocyte; Measures; Mediating; Messenger RNA; Mononuclear; Mus; Natural regeneration; Neutrophil Infiltration; Nucleic Acid Regulatory Sequences; Outcome; P-Selectin; P-selectin ligand protein; Pathogenesis; Pathology; Peroxidases; Polysaccharides; Prevention approach; Reporting; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Selectins; Smooth Muscle Myocytes; Staining method; Stains; Surface; T-Lymphocyte; Testing; Time; Transgenic Mice; arterial lesion; base; beta-1,3-Galactosyl-o-glycosyl-glycoprotein beta-1,6-N-acetylglucosaminyltransferase; expectation; feeding; glycosylation; injured; macrophage; monocyte; neutrophil; recombinase

DESCRIPTION: Lesions and neointima that forms following arterial injury are two of the major arterial pathologies in atherosclerosis. These two arterial pathologies are initiated by the recruitment of leukocytes to the arterial vessel walls. The long-term goal of this research is to determine the role of neutrophils in the genesis of arterial diseases. The central hypothesis of this proposal is that neutrophil recruitment to arterial vessel walls is required for the contribution of neutrophils to the development of arterial diseases. In contrast to the well-documented effect of mononuclear cells on the development of arterial diseases, especially spontaneous lesions, the involvement of neutrophils in these arterial diseases is disputable. However, one recent study has reported increased atherosclerosis in mice with a high neutrophil count and decreased atherosclerosis in mice treated with a neutrophil depletion antibody. Although that study points to an important role of neutrophils in atherosclerosis, the approaches it used alter neutrophil homeostasis, and it thus fails to establish a direct link between neutrophil recruitment to the arterial vessel wall and the development of arterial diseases under normal leukocyte homeostasis. Leukocyte recruitment to the arterial vessel wall is initiated by leukocyte tethering and rolling; the latter are mediated by leukocyte selectin ligands [PSGL-1 (ligand for P-selectin) and ligands for E- and L-selectin]. Glycosylation of selectin ligands is crucial for the binding of selectin ligands to the selectins. Core2 1-6-N- glucosaminyltransferase-I (C2GlcNAcT-I) is one of the enzymes that mediate the glycosylation of selectin ligands. Our previous studies have shown that knockout of C2GlcNAcT-I almost completely eliminates leukocyte recruitment to arterial vessel walls and suppresses the formation of spontaneous and neointimal lesions in mice. We have recently generated neutrophil-specific Cre transgenic mice, mice in which the expression of Cre recombinase and EGFP are driven by the regulatory region of the myeloperoxidase (MPO) gene (MPO-Cre mice). By breeding these MPO-Cre mice with floxed C2GlcNAcT-I mice, we have generated mice with C2GlcNAcT-I deficiency specifically in neutrophils. ApoE-/- mice lacking C2GlcNAcT-I specifically in neutrophils (Neu-C2GlcNAcT-I-/-/apoE-/- mice) were also generated. We have found that neutrophils in these mice fail to interact with the endothelium. The goal of this project is to use these mice to investigate the effect of neutrophil recruitment to arterial vessel walls on the formation of spontaneous atherosclerotic lesions and neointimal lesions. Specifically, in two arterial disease models we will examine whether neutrophil C2GlcNAcT- I deficiency affects the recruitment of monocytes and/or platelets to the arterial vessel wall, endothelial regeneration, the formation of spontaneous lesions and arterial neointima following arterial injury in apoE-/- mice. These studies will provide evidence that inhibition of neutrophil recruitment to arteries is a viable approach to the prevention and treatment of arterial diseases. PUBLIC HEALTH RELEVANCE: Results from the proposed research will demonstrate whether neutrophil recruitment to the arterial vessel wall is required for the contribution of neutrophils to the development of arterial diseases. The successful completion of this project will provide evidence for the inhibition of neutrophil homing to arterial vessel walls as one of the approaches for the prevention and treatment of arterial diseases.

动脉损伤后形成的病变和新生内膜是动脉粥样硬化的两种主要病理。这两种动脉病变是由白细胞聚集到动脉血管壁上引起的。这项研究的长期目标是确定中性粒细胞在动脉疾病发生中的作用。该建议的中心假设是中性粒细胞向动脉血管壁募集是中性粒细胞对动脉疾病发展的贡献所必需的。与单核细胞在动脉疾病，特别是自发性病变发展中的作用相比，中性粒细胞在这些动脉疾病中的参与是有争议的。然而，最近的一项研究报道，中性粒细胞计数高的小鼠动脉粥样硬化增加，而中性粒细胞消耗抗体治疗的小鼠动脉粥样硬化减少。尽管该研究指出了中性粒细胞在动脉粥样硬化中的重要作用，但其使用的方法改变了中性粒细胞的稳态，因此未能在正常白细胞稳态下建立中性粒细胞向动脉血管壁募集与动脉疾病发展之间的直接联系。白细胞聚集到动脉血管壁上是由白细胞系缚和滚动引起的；后者由白细胞选择素配体[PSGL-1 （p -选择素配体）和E-和l -选择素配体]介导。选择素配体的糖基化是选择素配体与选择素结合的关键。Core2 1-6-N-氨基葡萄糖转移酶（C2GlcNAcT-I）是介导选择素配体糖基化的酶之一。我们之前的研究表明，敲除C2GlcNAcT-I几乎完全消除了白细胞向动脉血管壁的募集，并抑制了小鼠自发性和新内膜病变的形成。我们最近培育了中性粒细胞特异性Cre转基因小鼠，在这些小鼠中，Cre重组酶和EGFP的表达是由髓过氧化物酶（MPO）基因的调控区域驱动的（MPO-Cre小鼠）。通过将这些MPO-Cre小鼠与固定的C2GlcNAcT-I小鼠杂交，我们产生了中性粒细胞特异性缺乏C2GlcNAcT-I的小鼠。在中性粒细胞中特异性缺乏C2GlcNAcT-I的ApoE-/-小鼠（new -C2GlcNAcT-I-/-/ ApoE-/-小鼠）也被生成。我们发现这些小鼠的中性粒细胞不能与内皮相互作用。本项目的目的是利用这些小鼠来研究中性粒细胞募集到动脉血管壁对自发性动脉粥样硬化病变和新内膜病变形成的影响。具体而言，在两种动脉疾病模型中，我们将研究中性粒细胞C2GlcNAcT- I缺乏是否影响apoE-/-小鼠动脉损伤后单核细胞和/或血小板向动脉血管壁的募集、内皮细胞再生、自发病变和动脉新内膜的形成。这些研究将提供证据，证明抑制中性粒细胞向动脉募集是预防和治疗动脉疾病的可行方法。