The role of syndecan-4 in the regulation of JNK activation in human neutrophils

syndecan-4 在调节人中性粒细胞 JNK 激活中的作用

基本信息

批准号：
7209928
负责人：
PATRICK G ARNDT
金额：
$ 26.16万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-02-09 至 2012-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7209928
关键词：
Acute Adaptor Signaling Protein Adherence Adhesions Area Binding Blocking Antibodies CXCR4 Receptors CXCR4 gene Cell Adhesion Cell Line Cell Surface Proteins Cell surface Cells Chemicals Condition Dependence Exposure to Extracellular Matrix Family Hela Cells Heparin Binding Heparin Lyase Heparitin Sulfate Human IL8 gene IL8RB gene Inflammation Inflammatory Inflammatory Response Interleukin-8B Receptor JUN gene Lead Ligands Lung Lung Inflammation MAPK14 gene Mediating Membrane Proteins Mitogen-Activated Protein Kinases Modeling Monocyte Chemoattractant Protein-1 Mus Neutrophil Infiltration Pathway interactions Phosphorylation Principal Investigator Process Protein Kinase C Protein Kinase C Alpha Protein Phosphatase 2A Regulatory Subunit PR53 Protein phosphatase Proteins Proteoglycan Regulation Research Personnel Role Scheme Side Signal Pathway Signal Transduction Small Interfering RNA Staging Techniques Up-Regulation chemokine chemokine receptor improved inhibitor/antagonist insight monocyte neutrophil novel presynaptic density protein 95 programs receptor stress-activated protein kinase 1 syndecan syndecan-4 zonula occludens-1 protein

项目摘要

DESCRIPTION (provided by applicant): The recruitment of monocytes to areas of acute inflammation is a later stage in the inflammatory process. We have previously shown that systemic inhibition of either p38 or c-Jun NH2-terminal kinase (JNK) limits neutrophil recruitment to the lung in an LPS-induced model of acute inflammation. As we will show, however, inhibition of JNK, but not p38, limits the pulmonary recruitment of monocytes, which occurs through monocyte chemoattractant protein 1 (MCP-1) and neutrophil dependent pathways. Understanding mechanisms that lead to JNK activation in neutrophils will, therefore, be fundamental to the understanding of monocyte recruitment. While activation of p38 in neutrophils occurs in suspended conditions, activation of JNK requires neutrophils to be cultured in adherent conditions, although the cell surface proteins that regulate cell adhesion, and thereby JNK activation, are poorly understood. One such family, the syndecans, bind to the extracellular matrix (ECM) inducing cell adhesion. In addition, syndecan-4 has been shown to co-associate with CXCR4, and we will show here with the IL-8 receptor CXCR2 in neutrophils. We propose to examine the role of syndecan-4 in regulating IL-8-induced JNK activation in neutrophils, and thereby expression of MCP-1, by examining if syndecan-4 enhances JNK activation via an upregulation of protein kinase C alpha (PKC() activity, examine mechanisms for the co-association of syndecan-4 with CXCR2, and the necessity of the co-association for the enhancement of IL-8-induced JNK activation. We will use several techniques including chemical inhibitors, blocking antibodies, small interfering RNAs, and protein transduction of neutrophils and the PLB-985 cell line, which can be differentiated into neutrophil-like cells. In addition, to confirm our findings from neutrophils and PLB-985 cells, we will utilize neutrophils derived from syndecan-4, CXCR2, and PKC( deficient mice. Utilizing the above techniques we will examine components that we hypothesize to regulate syndecan-4-induced PKC( and JNK activation, including PKC-( and PP2A, or to regulate the co-association of syndecan-4 with CXCR2, including the adaptor proteins CASK and syntenin. These studies will provide vital insights into mechanisms that regulate JNK activation in neutrophils and will suggest roles for novel pathways in the regulation of lung inflammation, and in particular pulmonary monocyte recruitment.

描述（由申请人提供）：单核细胞募集到急性炎症区域是炎症过程的后期。我们之前已经证明，在 LPS 诱导的急性炎症模型中，p38 或 c-Jun NH2 末端激酶 (JNK) 的全身抑制可限制中性粒细胞向肺部募集。然而，正如我们将要展示的，JNK 的抑制（而非 p38）限制了单核细胞的肺部募集，这是通过单核细胞趋化蛋白 1 (MCP-1) 和中性粒细胞依赖性途径发生的。因此，了解导致中性粒细胞 JNK 激活的机制对于了解单核细胞募集至关重要。虽然中性粒细胞中 p38 的激活发生在悬浮条件下，但 JNK 的激活需要在贴壁条件下培养中性粒细胞，尽管人们对调节细胞粘附以及由此调节 JNK 激活的细胞表面蛋白知之甚少。多配体家族就是这样的一个家族，它与细胞外基质（ECM）结合，诱导细胞粘附。此外，syndecan-4 已被证明与 CXCR4 共关联，我们在此将展示与中性粒细胞中的 IL-8 受体 CXCR2 的共关联。我们建议通过检查 syndecan-4 是否通过上调蛋白激酶 C α (PKC( ）活性，检查 syndecan-4 与 CXCR2 共关联的机制，以及共关联增强 IL-8 诱导的 JNK 激活的必要性。包括化学抑制剂、阻断抗体、小干扰 RNA 以及中性粒细胞和 PLB-985 细胞系的蛋白质转导，这些细胞可以分化为中性粒细胞样细胞。此外，为了证实我们在中性粒细胞和 PLB-985 细胞中的发现。将利用来自 syndecan-4、CXCR2 和 PKC（缺陷小鼠）的中性粒细胞。利用上述技术，我们将检查我们假设调节的成分syndecan-4 诱导 PKC( 和 JNK 激活，包括 PKC-( 和 PP2A)，或调节 syndecan-4 与 CXCR2 的共缔合，包括接头蛋白 CASK 和 syntenin。这些研究将为调节中性粒细胞中 JNK 激活的机制提供重要见解，并将提出新途径在调节肺部炎症，特别是肺单核细胞募集中的作用。