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Synovial mast cells in inflammatory arthritis

炎症性关节炎中的滑膜肥大细胞

基本信息

批准号：
8466273
负责人：
Richard L Stevens
金额：
$ 39.7万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-04-01 至 2014-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8466273
关键词：
Abbreviations Affect Animal Model Animals Arthritis Autoimmune Diseases Binding Bone Marrow Bovine Serum Albumin C57BL/6 Mouse Cartilage Cell Lineage Cells Chondrocytes Chromosomes Chymase Coculture Techniques Collection Complement 5a Complex Connective Tissue Deacetylase Degenerative polyarthritis Development Disease Enzyme-Linked Immunosorbent Assay Extracellular Matrix Family Fibroblasts Genes Genetic Transcription Green Fluorescent Proteins Heparin Human Hyaluronan Immature Mast Cell Immune In Vitro Inbred Mouse Inbreeding Inflammation Interleukins Joints Life Mass Spectrum Analysis Matrix Metalloproteinases Mediator of activation protein Mesenchymal Messenger RNA Modeling Molecular Mouse Strains Mus Orthologous Gene Pathway interactions Patients Peptide Hydrolases Phenotype Phosphate Buffer Physiology Platelet-Derived Growth Factor Play Polyacrylamide Gel Electrophoresis Polymerase Chain Reaction Progress Reports Proteins Proteoglycan Rattus Reagent Recombinants Research Rheumatoid Arthritis Role Saline Secretory Vesicles Serine Protease Sodium Dodecyl Sulfate-PAGE Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Staging Stem Cell Factor Synovial Fluid Synovial Membrane TNF gene Time Tissue Inhibitor of Metalloproteinases Tissues Transcript Transforming Growth Factors Transgenic Mice Transgenic Organisms Tryptase Tumor Necrosis Factor-alpha Untranslated Regions Work aggrecan cytokine gel electrophoresis in vivo inhibitor/antagonist insight joint injury mast cell mast cell protease 6 member mouse model novel public health relevance release factor serglycin sulfotransferase

项目摘要

DESCRIPTION (provided by applicant): The presence of elevated numbers of activated mast cells (MCs) in the joints of patients with rheumatoid arthritis (RA) raised the possibility in the 1980s that MCs participate in the inflammation, proteolytic damage, and/or remodeling of the arthritic joint. Using MC-deficient mice, we and others subsequently demonstrated that these immune cells do indeed contribute substantially to experimental inflammatory arthritis. Nevertheless, the relevant mediators produced by the Fc3RIII- and C5a-activated MCs in the synovium that adversely affect the joint had not been identified. Mouse MC protease (mMCP) 6 and mMCP-7 are members of the chromosome 17A3.3 family of tryptic proteases that are preferentially stored in the secretory granules of MCs as homotypic and heterotypic tetramers ionically bound to heparin-containing serglycin proteoglycans. The human ortholog of mMCP-6 is hTryptase-21. We generated enzymatically active mMCP-6, mMCP-7, and hTryptase-21. We also recently created inbred transgenic C57BL/6 mouse strains that differ in their expression of mMCP-6, mMCP-7, and heparin. Using the latter mice, we showed that C57BL/6 mice that lack tryptase7heparin complexes have reduced inflammation and diminished loss of aggrecan proteoglycans from their cartilage in two different mouse models of inflammatory arthritis. The overall objectives of the proposed studies are to use complementary approaches to determine at the molecular level the roles of mouse and human tetramer-forming tryptases in RA and mouse models of this autoimmune disorder. Our genetically modified mouse strains, recombinant tryptases, and other key protease-specific reagents will be used in the proposed studies to determine how MC-restricted tetramer-forming human and mouse tryptases affect fibroblast-like synoviocytes (FLS) and chondrocytes at multiple levels in varied in vitro and in vivo studies, as well as how these mesenchymal cells affect MCs. Additional studies will be carried out to determine how exocytosed mMCP-6 and hTryptase-21 are negatively regulated in the arthritic joint, and even in MCs themselves. The factors and mechanisms that control the expression of enzymatically active tryptases inside MCs have not been identified. To this end, we recently discovered that FLS-derived IL-33 induces MCs to increase their expression of tryptases at the mRNA and protein levels. Thus, additional mechanistic studies are proposed to understand how IL-33 controls the transcription of the mMCP-6 and hTryptase-21 genes in MCs and/or the stability of their transcripts. In combination, these studies promise to provide novel insights into both MC and synovial physiology, as well as elucidate novel disease mechanisms in human inflammatory arthritis.

描述（由申请人提供）：20世纪80年代，类风湿性关节炎（RA）患者关节中活化肥大细胞（MC）数量增加，这增加了MC参与关节炎关节炎症、蛋白水解损伤和/或重塑的可能性。使用MC缺陷小鼠，我们和其他人随后证明，这些免疫细胞确实对实验性炎症性关节炎有很大贡献。然而，尚未确定滑膜中Fc 3RIII和C5 a激活的MC产生的对关节产生不利影响的相关介质。小鼠MC蛋白酶（mMCP）6和mMCP-7是胰蛋白酶的染色体17A3.3家族的成员，其优先作为与含肝素的丝甘蛋白蛋白聚糖离子结合的同型和异型四聚体储存在MC的分泌颗粒中。mMCP-6的人类直系同源物是hTryptase-21。我们产生了具有酶活性的mMCP-6、mMCP-7和hTryptase-21。我们最近还建立了近交系转基因C57 BL/6小鼠品系，其mMCP-6、mMCP-7和肝素的表达不同。使用后一种小鼠，我们发现缺乏类胰蛋白酶7肝素复合物的C57 BL/6小鼠在两种不同的炎症性关节炎小鼠模型中减少了炎症和软骨中聚集蛋白聚糖蛋白聚糖的损失。拟议的研究的总体目标是使用互补的方法，以确定在分子水平上的作用，小鼠和人类四聚体形成类胰蛋白酶在RA和小鼠模型的这种自身免疫性疾病。我们的转基因小鼠品系，重组胰蛋白酶和其他关键的蛋白酶特异性试剂将用于拟议的研究，以确定MC限制性四聚体形成人类和小鼠胰蛋白酶如何影响成纤维细胞样滑膜细胞（FLS）和软骨细胞在多个水平在不同的体外和体内研究，以及这些间充质细胞如何影响MC。将进行进一步的研究，以确定外切mMCP-6和hTryptase-21是如何在关节炎关节，甚至在MC本身的负调控。控制MC内酶活性胰蛋白酶表达的因素和机制尚未确定。为此，我们最近发现，FLS衍生的IL-33诱导MC在mRNA和蛋白水平上增加其类胰蛋白酶的表达。因此，提出了另外的机制研究以理解IL-33如何控制MC中mMCP-6和hTryptase-21基因的转录和/或其转录物的稳定性。结合起来，这些研究有望为MC和滑膜生理学提供新的见解，并阐明人类炎症性关节炎的新疾病机制。