Annexins and Osteoarthritis

膜联蛋白和骨关节炎

基本信息

批准号：
9495667
负责人：
THORSTEN KIRSCH
金额：
$ 37.29万
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：
2014
资助国家：
美国
起止时间：
2014-06-01 至 2020-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9495667
关键词：
Affect Aging Annexin A6 Annexins Binding Biological Assay Calpain Cardiovascular Diseases Cartilage Cell membrane Cells Chondrocytes Complex Cysteine Proteinase Inhibitors Degenerative polyarthritis Dependence Development Diabetes Mellitus Disease Event Goals Human Individual Injections Interleukin-1 Interleukin-1 beta Joints Knee joint Knock-out Knockout Mice Lead Ligaments Malignant Neoplasms Medial Mediating Membrane Meniscus structure of joint Modeling Mus Nature Operative Surgical Procedures Pathology Pathway interactions Phase Phenotype Process Proteins Publishing Recombinants Regulation Reporter Research Role Signal Pathway Signal Transduction Solid Surface Plasmon Resonance Synovial Membrane TNFSF11 gene Tendon structure Testing Therapeutic Transfection Tumor necrosis factor receptor 11b annexin A5 articular cartilage base beta catenin bone loss calpastatin collateral ligament cytokine experimental study insight mutant new therapeutic target novel osteoclastogenesis overexpression p65 public health relevance skeletal subchondral bone therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Annexins became attractive therapeutic targets for the treatment of various diseases, including cancer, cardiovascular diseases and diabetes, because of their ability to modulate the activities of important signaling pathways involved in disease pathology. Annexin (Anx) A5 and AnxA6 are highly expressed in osteoarthritic (OA) cartilage; however, nothing is known about their role in OA pathology. Two major signaling pathways with roles in OA are NF-κB and the Wnt/ß-catenin (Wnt), encouraging us to examine the potential interaction between AnxA5 and AnxA6 and these signaling pathways. NF-κB signaling pathway, which is being activated in articular chondrocytes by various cytokines, including interleukin (IL)-1, is one of the major catabolic signaling pathways in OA, whereas the Wnt signaling pathway has been recently shown to act anti-catabolically in human articular chondrocytes while acting catabolically in mouse chondrocytes. Our preliminary findings show that AnxA5 and AnxA6 stimulate NF-κB signaling while inhibiting Wnt signaling. In addition, our findings suggest that AnxA5 modulates NF-κB and Wnt signaling via direct interaction with calpastatin, an inhibitor of the cysteine protease calpain, thereby stimulating calpain activity. Calpain stimulates NF-κB signaling, while inhibiting Wnt signaling. AnxA6, on the other hand, stimulates NF-κB signaling via direct binding to the p65 unit of the NF-κB complex, while it inhibits Wnt signaling via interfering with membrane association of the Wnt signaling complex, which is required for Wnt signaling activation. Based on these findings, we hypothesize that AnxA5 and AnxA6 act via different mechanisms, to modulate NF-κB and Wnt signaling to stimulate cartilage destruction during OA pathology. To test our hypothesis, we are proposing two aims. In the first aim we will determine the nature of the AnxA5/calpastatin and AnxA6/p65 interactions, and how these interactions affect NF-κB signaling activity. In addition, we will determine how the AnxA5/calpastatin interaction affect Wnt signaling and how Ca2+-dependent plasma membrane association of AnxA6 interferes with the membrane association of the Wnt signaling complex and ultimately Wnt signaling activity. Furthermore, we will determine how AnxA5 and AnxA6 individually and together affect NF-κB and Wnt signaling during aging, IL-1 injection in the knee joint or surgically induced OA in AnxA5 and AnxA6 single and double knockout mice. In Aim 2, we will determine the effect of annexin-mediated modulation of these signaling pathways on the function and phenotype of human articular chondrocytes. Finally, we will determine how annexin-mediated inhibition of Wnt signaling affects OPG expression in AnxA5 and AnxA6 single and double knockout articular chondrocytes and AnxA5 and AnxA6 overexpressing chondrocytes and ultimately osteoclastogenesis and subchondral bone changes in OA pathology. We expect that the successful completion of this proposal will provide novel mechanisms stimulating cartilage destruction during OA pathology and novel therapeutic targets for the treatment of OA.

描述（由适用提供）：膜联蛋白成为治疗各种疾病（包括癌症，心血管疾病和糖尿病）的有吸引力的治疗靶标，因为它们有能力调节涉及疾病病理学的重要信号传导途径的活性。 Annexin（Anx）A5和AnxA6在骨关节炎（OA）软骨中高度表达；但是，他们在OA病理学中的作用一无所知。在OA中具有作用的两个主要信号通路是NF-κB和Wnt/ß-catenin（Wnt），鼓励我们检查Anxa5和Anxa6以及这些信号通路之间的潜在相互作用。 NF-κB信号传导途径在包括白介素（IL）-1在内的各种细胞因子在关节软骨细胞中激活的NF-κB信号通路是OA中的主要分解代谢信号通路之一，而Wnt信号通路最近已显示出在人类动臂旁核心的抗catibolational catibolational catodrocy catborcy catbondocy catbodolation catborcy catborcy catborcy catborcy catborcy。我们的初步发现表明，AnxA5和AnxA6刺激NF-κB信号传导，同时抑制WNT信号传导。此外，我们的发现表明，AnxA5通过与Calpastatin直接相互作用（半胱氨酸蛋白钙蛋白酶的抑制剂）通过直接相互作用来调节NF-κB和Wnt信号传导，从而刺激钙蛋白酶活性。钙蛋白酶刺激NF-κB信号传导，同时抑制Wnt信号传导。另一方面，AnxA6通过直接结合NF-κB复合物的p65单位刺激NF-κB信号传导，而它通过干扰Wnt信号传导复合物的膜关联来抑制Wnt信号传导，这是Wnt信号激活所必需的。根据这些发现，我们假设Anxa5和Anxa6通过不同的机制起作用，以调节NF-κB和Wnt信号传导以刺激OA病理期间的软骨破坏。为了检验我们的假设，我们提出了两个目标。在第一个目标中，我们将确定Anxa5/calpastatin和Anxa6/p65相互作用的性质，以及这些相互作用如何影响NF-κB信号传导活性。此外，我们将确定Anxa5/calpastatin的相互作用如何影响Wnt信号传导，以及CA2+依赖性的质膜如何与Wnt信号传导复合物和Wnt信号活性的膜关联与膜关联的质膜关联。此外，我们将确定Anxa5和AnxA6在衰老期间如何单独影响NF-κB和Wnt信号，膝关节中的IL-1注射或Anxa5中的手术诱导OA，而Anxa6单一和双敲除小鼠。在AIM 2中，我们将确定膜联蛋白介导的这些信号通路对人关节软骨细胞的功能和表型的作用。最后，我们将确定膜联蛋白介导的WNT信号传导如何影响AnxA5和AnxA6单一和双基因敲除关节软骨细胞和ANXA5和ANXA6中的OPG表达，并过表达软骨细胞，并最终使OA病理学中的骨质肿瘤发生和骨内部骨变化。我们预计，该提案的成功完成将提供新的机制，以刺激OA病理学期间的软骨破坏和用于治疗OA的新型治疗靶标。