Pathogenesis of Inflammation-driven Intervertebral Disc Herniation: The Role of Syndecan 4

炎症驱动的椎间盘突出症的发病机制：Syndecan 4 的作用

基本信息

批准号：
9754682
负责人：
Makarand V Risbud
金额：
$ 46.25万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9754682
关键词：
ADAMTS Address American Animal Model Animals Apoptosis Back Back Pain Biomechanics CXCL12 gene Catabolism Cell Aging Cell surface Cells Chronic Collagen Defect Disease Etiology Fibronectins Foundations Future Genes Goals Heparan Sulfate Proteoglycan Heparitin Sulfate Human IL8 gene Immune Immune Cell Activation Incidence Individual Infiltration Inflammation Inflammatory Interleukin-1 beta Intervention Intervertebral disc structure Investigation Lead Learning Link Low Back Pain MMP3 gene Macrophage Activation Magnetic Resonance Imaging Matrix Metalloproteinases Measures Mediating Mediator of activation protein Molecular Molecular Structure Motion Mus Neck Pain Occupations Outcome Study Pain Pathogenesis Pathway interactions Phase Play Process Production Property Proteoglycan RANTES Regulation Role Side Signal Transduction Slipped Disk Solid Stromal Cell-Derived Factor 1 Stromelysin 1 Structure TNF gene Testing Time Tissues Transgenic Mice Trauma Unspecified or Sulfate Ion Sulfates Vertebral column Work aggrecan aggrecanase cell motility chemokine cytokine disability discogenic pain experience human tissue in vivo inflammatory milieu intervertebral disk degeneration loss of function macrophage migration mutant new therapeutic target novel nucleus pulposus overexpression prevent protein degradation stem syndecan-4 syndecan4 synergism therapy development transcriptome sequencing

项目摘要

The relationship between the etiology of intervertebral disc degeneration and low back pain has been subjected to considerable scrutiny. Degeneration is thought to be initiated by the abnormal production of a number of pro-inflammatory cytokines, in particular TNF-α and IL-1β. While these studies have shown involvement of cytokines in pathogenesis of disc degeneration and herniation, progress has been slow in delineating new therapeutic targets to block inflammation. This deficit stems from our lack of understanding of molecular mediators that regulate and link cytokine-dependent matrix degradation with immune cell migration into the disc through annular defects. Interestingly, our ongoing work has found an unexpected synergism between heparan sulfate (HS) proteoglycan syndecan4 (SDC4) and these two key inflammatory cytokines. The goal of the proposed investigations is to build on the central role of SDC4 in the pathogenesis of the disc herniation and inflammation. We have formulated two interrelated hypotheses each of which addresses a separate but linked phase of the disease process. The first phase relates to the effect of the inflammatory cytokines on the regulation of aggrecan rich matrix degradation. We will test the hypothesis that SDC4 through HS side chains promotes cytokine-dependent aggrecan rich matrix degradation through ADAMTS and MMP activation. Using gain and loss-of-function studies of SDC4 in NP cells treated with cytokines and RNA-Seq approach we will identify novel SDC4 responsive genes. We will also measure the contribution of MMPs, ADAMTS-1 and ADAM17 produced by NP cells in SDC4 shedding. In addition, we will determine how changes in SDC4 levels relate to aggrecan and collagen turnover in human tissues. The second stage of the disease process is characterized by structural changes in the NP and AF and formation of annular tears and herniations. We propose to test the hypothesis that that in inflammatory milieu of the herniated disc, SDC4 plays an important role in macrophage activation and migration by controlling the activity of select chemokines (CCL5, IL-8 and SDF-1) secreted by the disc cells. This will be achieved using primary macrophages isolated from SDC4-/- mice. Using well characterized painful human disc tissues, we will link changes in chemokines and SDC4 levels to immune cell activation, infiltration and matrix degradation. Finally, to explore the in vivo importance of SDC4 in pathogenesis of TNF--driven disc herniation, we will cross hTNFtg mice with SDC4 -/- animals to generate hTNFtg,SDC4-/- mice. To establish if a deficiency in SDC4 provides protection against cytokine-dependent disc herniation, we will compare molecular, structural and biomechanical properties of the motion segments with that of hTNFtg. To our knowledge, this would be the first attempt to address the contribution of SDC4 to mechanisms linking initiation and propagation of the degenerative cascade and herniation driven by inflammatory cytokines. These studies will provide a foundation for future development of interventional strategies to target SDC4 dependent catabolic and inflammatory phases of degenerative disc disease.

椎间盘变性的病因与腰痛之间的关系已受到进行进行大量审查。堕胎被认为是通过异常产生的许多促炎性细胞因子，特别是TNF-α和IL-1β。尽管这些研究表明椎间盘变性和催眠的发病机理中的细胞因子，描述新的进展缓慢治疗靶标可阻止炎症。由于我们对分子缺乏了解，这种赤字步骤调节和连接细胞因子依赖性基质降解与免疫细胞迁移到该的介体通过环形缺陷盘。有趣的是，我们正在进行的工作发现硫酸肝素（HS）蛋白聚糖Syndecan4（SDC4）和这两种关键的炎症细胞因子。目标拟议的调查是建立在SDC4在椎间盘突出的发病机理中的核心作用的基础和炎症。我们已经提出了两个相互关联的假设，每个假设都解决了一个单独的假设疾病过程的联系阶段。第一阶段涉及炎症细胞因子对调节富集蛋白矩阵降解的调节。我们将测试SDC4通过HS侧链的假设通过ADAMTS和MMP激活促进依赖细胞因子的富集蛋白矩阵降解。使用在用细胞因子和RNA-seq方法处理的NP细胞中SDC4的功能丧失研究我们将识别新型的SDC4响应基因。我们还将衡量MMP，ADAMTS-1和 NP细胞在SDC4脱落中产生的ADAM17。此外，我们将确定SDC4级别的变化与人体组织中的摄取和胶原蛋白更新有关。疾病过程的第二阶段是其特征是NP和AF的结构变化以及环形撕裂和疝的形成。我们提议检验以下假设，即在椎间盘突出的炎症环境中，SDC4发挥着重要的作用通过控制选择趋化因子的活性（CCL5，IL-8和 SDF-1）由椎间盘细胞分泌。使用从SDC4 - / - 小鼠中分离出来的主要巨噬细胞来实现这一点。使用良好特征的疼痛人盘组织，我们将将趋化因子和SDC4水平的变化联系起来免疫细胞激活，浸润和基质降解。最后，探索体内SDC4的重要性在TNF-驱动的椎间盘突出症的发病机理中，我们将用SDC4 - / - 动物穿过HTNFTG小鼠 htnftg，sdc4 - / - 小鼠。确定SDC4缺陷是否提供了针对细胞因子依赖性椎间盘的保护疝气，我们将比较运动节段的分子，结构和生物力学特性 htnftg的。据我们所知，这将是解决SDC4对连接了主动性和繁殖级联的机制炎症细胞因子。这些研究将为未来介入的发展提供基础针对退化性椎间盘疾病的依赖性分解代谢和炎症阶段的策略。