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的有吸引力的治疗靶点，包括癌症， 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AnxA 6在骨关节炎中高表达 (OA)软骨;然而，关于它们在OA病理学中的作用一无所知。两种主要的信号通路 在OA中起作用的是NF-B和Wnt/-连环蛋白（Wnt），这促使我们研究潜在的相互作用 和这些信号通路之间的联系。NF-B信号通路被激活 在关节软骨细胞中由各种细胞因子，包括白细胞介素（IL）-1，是主要的分解代谢之一， 信号通路，而Wnt信号通路最近已被证明具有抗分解代谢作用 在人关节软骨细胞中起作用，而在小鼠软骨细胞中起分解代谢作用。我们的初步研究结果 显示AnxA 5和AnxA 6刺激NF-B信号传导，同时抑制Wnt信号传导。此外，我们的发现 提示AnxA 5通过与钙蛋白酶抑制剂直接相互作用调节NF-B和Wnt信号传导，钙蛋白酶抑制剂是一种 半胱氨酸蛋白酶钙蛋白酶，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，AnxA 6通过直接结合p65单位刺激NF-B信号传导 NF-B复合物，而它通过干扰Wnt的膜缔合来抑制Wnt信号传导。 信号复合物，其是Wnt信号激活所需的。基于这些发现，我们假设 AnxA 5和AnxA 6通过不同的机制来调节NF-B和Wnt信号， OA病理过程中的软骨破坏。为了验证我们的假设，我们提出了两个目标。上 我们的目的是确定AnxA 5/calpastatin和AnxA 6/p65相互作用的性质，以及这些相互作用是如何发生的。 相互作用影响NF-B信号传导活性。此外，我们还将确定AnxA 5/钙蛋白酶抑制蛋白相互作用如何影响 影响Wnt信号传导以及AnxA 6的Ca 2+依赖性质膜结合如何干扰Wnt信号传导。 Wnt信号传导复合物的膜结合和最终的Wnt信号传导活性。此外，我们将 确定AnxA 5和AnxA 6如何单独和共同影响衰老过程中NF-B和Wnt信号传导，IL-1 在AnxA 5和AnxA 6单和双敲除小鼠中，膝关节注射或手术诱导的OA。在 目的2，我们将确定膜联蛋白介导的这些信号通路的调节对功能的影响， 和人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 Wnt信号对AnxA 5和AnxA 6单、双敲除关节软骨细胞OPG表达的影响 以及AnxA 5和AnxA 6过表达的软骨细胞，并最终导致破骨细胞生成和软骨下骨 OA病理学变化。我们预计，该提案的成功完成将提供新的 在OA病理过程中刺激软骨破坏的机制和新的治疗靶点， 治疗OA！