Regulation of chondrocytes by extracellular matrix protein.

细胞外基质蛋白对软骨细胞的调节。

基本信息

批准号：
8431781
负责人：
Steven B Abramson
金额：
$ 34.43万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-01 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8431781
关键词：
Address Adult Affect Age Area Binding Proteins Biological Process Bone Development Cartilage Cartilage Diseases Cell Culture Techniques Cell Surface Receptors Cells Characteristics Chondrocytes Collagen Collagen Type II Complementary DNA Complex Degenerative polyarthritis Deterioration Dinoprostone Disease Disease Progression Embryo Epiphysial cartilage Event Extracellular Matrix Proteins Gene Expression Genes Growth Histologic Human Hybrids Hypertrophy Immunoprecipitation In Vitro Joints Knockout Mice Lead Length Lesion Limb structure Link Matrix Metalloproteinases Medial Mediating Metabolism Modeling Molecular Mus Neurons Operative Surgical Procedures Osteoarthrosis Deformans Osteocalcin Osteogenesis Pathogenesis Pathway interactions Peptide Hydrolases Peptides Phenotype Plasmin Play Population Process Property Protein Family Proteins Proteomics Public Health Regulation Relative (related person)Research Proposals Rodent Role Skeletal Development Specimen Subgroup Technology Testing Tissues Transgenic Organisms Yeasts abstracting articular cartilage base cartilage repair collagenase 3 design disability improved in vivo insight knockout gene loss of function mature animal member mineralization mouse model novel novel strategies overexpression premature prevent protein protein interaction receptor research study tibia yeast two hybrid system

项目摘要

Abstract. F-spondin is a member of a family of proteins that collectively belong to a subgroup of TSR (thrombospondin) type I class molecules. We have discovered that F-spondin expression is significantly increased in osteoarthritic cartilage as well as in rodent meniscectomy models of OA. Preliminary studies indicate that F-spondin has significant effects on human chondrocyte metabolism and is also expressed in the hypertrophic regions of embryonic growth plates where it acts to regulate mineralization and endochondral bone formation. This proposal is designed to characterize the functional effects of F-spondin, which have major and previously unrecognized implications for the progression of OA. We will test two central hypotheses: 1) F-spondin modulates collagen degradation via unrecognized pathways that include activation of TGF-¿ and induction of MMPs and 2) F-spondin induces hypertrophic differentiation of articular chondrocytes and plays an essential role in the regulation of mineralization and endochondral bone formation. The specific aims designed to test these hypotheses are as follows: In SPECIFIC AIM 1 we will investigate the effects of F-spondin on the regulation of hypertrophy and mineralizing activity in human articular chondrocytes in vitro culture models. Cartilage specimens will be examined histologically to investigate the link between F-spondin and other characteristic hypertrophic/ossification markers of OA chondrocytes. In SPECIFIC AIM 2 we will investigate the molecular mechanism(s) of F-spondin-mediated collagen degradation in OA cartilage. Explant or cell cultures will be used to a) identify MMPs induced by F- spondin and examine their role in F-spondin-mediated collagen degradation b) establish the role of TGF-¿ in modulation of F-spondin functions and c) compare functional activity of the full length F-spondin molecule relative to its proteolytic fragments. In SPECIFIC AIM 3 we will identify and characterize the interacting proteins of F-spondin. a) investigate the interaction of F-spondin with the Latency-associated peptide (LAP) of the latent TGF-¿ complex and b) utilize yeast 2 hybrid and proteomic technologies to identify novel F- spondin binding proteins (proteases, receptors, matrix molecules) that may regulate its activity in articular cartilage. In SPECIFIC AIM 4 we will investigate the expression and function of F-spondin in cartilage in vivo. We will investigate F-spondin expression in cartilage in vivo i) during endochondral bone development and ii) in the mouse meniscectomy model of OA. We will generate an F-spondin knockout mouse and characterize the changes in cartilage phenotype during endochondral bone development and OA disease progression. Understanding the regulation of chondrocyte functions by F-spondin could lead to novel strategies for cartilage repair and disease modifying treatments for osteoarthritis.

摘要：F-spondin 是一个蛋白质家族的成员，这些蛋白质共同属于 TSR 的一个亚组。 (thrombospondin) I 类分子我们发现 F-spondin 表达显着。骨关节炎软骨以及 OA 啮齿动物半月板切除模型中的增加。表明F-spondin对人类软骨细胞代谢有显着影响，并且也表达于胚胎生长板的肥大区域，其作用是调节矿化和该提案旨在表征 F-spondin 的功能作用，这对 OA 的进展具有重大且之前未被认识到的影响，我们将测试其中两个。中心假设：1) F-spondin 通过未知途径调节胶原蛋白降解，包括 TGF-¿的激活和 MMP 的诱导，2) F-spondin 诱导关节的肥大分化软骨细胞在矿化和软骨内骨的调节中起着重要作用旨在检验这些假设的具体目标如下：在具体目标 1 中，我们将研究 F-spondin 对人体肥大和矿化活性调节的影响关节软骨细胞体外培养模型将进行组织学检查。研究 F-spondin 与 OA 其他特征性肥厚/骨化标志物之间的联系在 SPECIFIC AIM 2 中，我们将研究 F-spondin 介导的分子机制。 OA 软骨中的胶原蛋白降解将用于 a) 鉴定 F- 诱导的 MMP。 spondin 并检查其在 F-spondin 介导的胶原蛋白降解中的作用 b) 确定 TGF-¿ c) 比较全长 F-spondin 分子的功能活性相对于其蛋白水解片段，在 SPECIFIC AIM 3 中，我们将识别并表征相互作用。 F-spondin 蛋白 a) 研究 F-spondin 与潜伏相关肽 (LAP) 的相互作用。潜在 TGF-¿ b) 利用酵母 2 杂交和蛋白质组技术来鉴定新型 F- spondin 结合蛋白（蛋白酶、受体、基质分子）可调节其在关节中的活性在 SPECIFIC AIM 4 中，我们将研究 F-spondin 在软骨中的表达和功能。我们将研究软骨内骨发育过程中的 F-spondin 体内表达。 ii) 在 OA 的小鼠半月板切除模型中，我们将生成 F-spondin 敲除小鼠，并且表征软骨内骨发育和 OA 疾病期间软骨表型的变化了解 F-spondin 对软骨细胞进展功能的调节可能会带来新的发现软骨修复和骨关节炎疾病改变治疗的策略。