Annexins and Osteoarthritis

膜联蛋白和骨关节炎

• 批准号: 9305225
• 负责人: THORSTEN KIRSCH
• 金额: $ 6.74万
• 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/9305225
• 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; Health; Human; Individual; Injection of therapeutic agent; 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; 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; bone loss; calpastatin; collateral ligament; cytokine; insight; mutant; new therapeutic target; novel; osteoclastogenesis; overexpression; p65; research study; skeletal; therapeutic target

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（由申请人提供）：膜联蛋白成为治疗各种疾病的有吸引力的治疗靶标，包括癌症、心血管疾病和糖尿病，因为它们能够调节疾病病理学中涉及的重要信号传导途径的活性。膜联蛋白 (Anx) A5 和 AnxA6 在骨关节炎 (OA) 软骨中高表达；然而，它们在 OA 病理学中的作用尚不清楚。在 OA 中发挥作用的两条主要信号通路是 NF-κB 和 Wnt/ß-连环蛋白 (Wnt)，这鼓励我们研究 AnxA5 和 AnxA6 与这些信号通路之间的潜在相互作用。 NF-κB信号通路在关节软骨细胞中被多种细胞因子（包括白细胞介素（IL）-1）激活，是OA中主要的分解代谢信号通路之一，而Wnt信号通路最近已被证明在人关节软骨细胞中具有抗分解代谢作用，而在小鼠软骨细胞中具有分解代谢作用。我们的初步研究结果表明，AnxA5 和 AnxA6 刺激 NF-κB 信号传导，同时抑制 Wnt 信号传导。此外，我们的研究结果表明，AnxA5 通过与半胱氨酸蛋白酶钙蛋白酶抑制剂 Calpastatin 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。 Calpain 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 单位来刺激 NF-κB 信号传导，同时通过干扰 Wnt 信号传导复合物的膜关联来抑制 Wnt 信号传导，而 Wnt 信号传导复合物是 Wnt 信号传导激活所必需的。基于这些发现，我们假设 AnxA5 和 AnxA6 通过不同的机制发挥作用，调节 NF-κB 和 Wnt 信号传导，从而在 OA 病理过程中刺激软骨破坏。为了检验我们的假设，我们提出两个目标。第一个目标是确定 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 相互作用如何影响 Wnt 信号传导，以及 AnxA6 的 Ca2+ 依赖性质膜关联如何干扰 Wnt 信号复合物的膜关联以及最终的 Wnt 信号传导活性。此外，我们将确定 AnxA5 和 AnxA6 在老化、膝关节注射 IL-1 或手术诱导 AnxA5 和 AnxA6 单敲除和双敲除小鼠中的 OA 过程中如何单独和共同影响 NF-κB 和 Wnt 信号传导。在目标 2 中，我们将确定膜联蛋白介导的这些信号传导途径的调节对人类关节软骨细胞的功能和表型的影响。最后，我们将确定膜联蛋白介导的 Wnt 信号传导抑制如何影响 AnxA5 和 AnxA6 单敲除和双敲除关节软骨细胞以及 AnxA5 和 AnxA6 过表达软骨细胞中的 OPG 表达，并最终影响 OA 病理学中的破骨细胞生成和软骨下骨变化。我们期望该提案的成功完成将为 OA 病理过程中刺激软骨破坏的新机制和治疗 OA 的新治疗靶点提供新的机制。