Annexins and Osteoarthritis

膜联蛋白和骨关节炎

• 批准号: 8629242
• 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
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8629242
• 关键词: Affect; Aging; Annexin A6; Annexins; Binding; Biological Assay; Breeding; Calpain; Cardiovascular Diseases; Cartilage; Cell membrane; Cells; Chondrocytes; Complex; Cysteine Proteinase Inhibitors; Degenerative polyarthritis; Dependency; Development; Diabetes Mellitus; Disease; Event; Goals; Human; Individual; Injection of therapeutic agent; Interleukin-1; Joints; Knee joint; Knock-out; Knockout Mice; Lead; Ligaments; Malignant Neoplasms; Medial; Mediating; Membrane; Meniscus structure of joint; Modeling; Mus; Nature; Osteoarthrosis Deformans; 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; bone; bone loss; calpastatin; collateral ligament; cytokine; insight; mutant; new therapeutic target; novel; osteoclastogenesis; overexpression; p65; public health relevance; research study; skeletal; therapeutic target

Project Summary 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-¿B and the Wnt/¿-catenin (Wnt), encouraging us to examine the potential interaction between AnxA5 and AnxA6 and these signaling pathways. NF-¿B signaling pathway, which is being activated in articular chondrocytes by various cytokines, including interleukin (IL)-1, is one of the major catabolic signaling pathway 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-¿B signaling while inhibiting Wnt signaling. In addition, our findings suggest that AnxA5 modulates NF-¿B and Wnt signaling via direct interaction with calpastatin, an inhibitor of the cysteine protease calpain, thereby stimulating calpain activity. Calpain stimulates NF-¿B signaling, while inhibiting Wnt signaling. AnxA6, on the other hand, stimulates NF-¿B signaling via direct binding to the p65 unit of the NF-¿B 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-¿B 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-¿B 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-¿B 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.!

项目摘要 膜联蛋白成为治疗各种疾病（包括癌症）的有吸引力的治疗靶标 心血管疾病和糖尿病，因为它们能够调节重要信号的活动 参与疾病病理学的途径。 Annexin（Anx）A5和Anxa6在骨关节炎中高度表达 （OA）软骨；但是，他们在OA病理学中的作用一无所知。两个主要的信号通路与 OA中的角色是NF -€和Wnt/®-Catenin（Wnt），鼓励我们检查潜在的相互作用 在Anxa5和AnxA6之间以及这些信号通路之间。 NF-€b信号通路，正在激活 在包括白介素（IL）-1在内的各种细胞因子的关节软骨细胞中，是主要分解代谢之一 OA中的信号通路，而Wnt信号通路最近已显示出抗代谢作用 在人类关节软骨细胞中，同时在小鼠软骨细胞中作用。我们的初步发现 表明AnxA5和AnxA6在抑制Wnt信号传导的同时刺激NF- - B信号传导。此外，我们的发现 建议Anxa5通过与Calpastatin直接相互作用来调节NF- - B和Wnt信号传导 半胱氨酸蛋白钙蛋白酶，从而刺激钙蛋白酶活性。钙蛋白酶刺激nf-€b信号，而 抑制Wnt信号传导。另一方面，AnxA6通过直接结合p65单位刺激NF-€b信号传导 NF-€b复合物的，而通过干扰Wnt的膜关联，它抑制Wnt信号传导 信号复合物，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-€和Wnt信号传导以刺激 OA病理学期间的软骨破坏。为了检验我们的假设，我们提出了两个目标。在第一个 目的，我们将确定Anxa5/calpastatin和Anxa6/p65相互作用的性质，以及如何 相互作用会影响NF- - B信号活性。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+依赖性质膜如何干扰 Wnt信号复合物和Wnt信号活性的膜关联。此外，我们会的 确定Anxa5和Anxa6在衰老过程中如何单独影响NF-®B和Wnt信号传导 在膝关节或外科手术诱导的OA中注射Anxa5和Anxa6单敲除小鼠。在 AIM 2，我们将确定这些信号途径的膜联蛋白介导的调节对功能的影响 和人类关节软骨细胞的表型。最后，我们将确定膜联蛋白介导的抑制作用 Wnt信号会影响Anxa5和AnxA6单敲门和双基因敲除关节软骨细胞中的OPG表达 以及Anxa5和AnxA6过表达软骨细胞，最终的破骨细胞发生和软骨下骨 OA病理学的变化。我们预计该提案的成功完成将提供新颖 在OA病理学期间刺激软骨破坏的机制和新的治疗靶标的机制 OA的处理！