Annexins and Osteoarthritis

膜联蛋白和骨关节炎

基本信息

批准号：
9272836
负责人：
THORSTEN KIRSCH
金额：
$ 38.64万
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 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9272836
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 Dependency Development Diabetes Mellitus Disease Event Goals 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 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，因为它们能够调节与疾病病理学相关的重要信号传导途径的活性。 AnxA6 在骨关节炎 (OA) 软骨中高表达；然而，对于它们在 OA 病理学中的作用尚不清楚，其中两个主要信号通路是 NF-κB 和 OA。 Wnt/ß-连环蛋白 (Wnt)，鼓励我们检查 AnxA5 和 AnxA6 与这些信号通路之间的潜在相互作用，该信号通路在关节软骨细胞中被各种细胞因子（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类关节软骨细胞中具有抗分解代谢作用，而在小鼠中则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，钙蛋白酶刺激 NF-κB 信号传导，同时抑制 AnxA6 信号传导。 NF-κB 信号传导通过直接结合 NF-κB 复合物的 p65 单元，同时它通过干扰 Wnt 信号传导复合物的膜关联来抑制 Wnt 信号传导，而 Wnt 信号传导复合物是 Wnt 信号传导激活所必需的。 AnxA5 和 AnxA6 通过不同的机制发挥作用，在 OA 病理过程中调节 NF-κB 和 Wnt 信号传导以刺激软骨破坏。为了检验我们的假设，我们提出了两个目标。 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。与 Wnt 信号复合物的膜关联以及最终的 Wnt 信号活性此外，我们将确定 AnxA5 和 AnxA6 如何单独和共同影响 NF-κB 和AnxA5 和 AnxA6 单敲除和双敲除小鼠衰老过程中的 Wnt 信号传导、膝关节注射 IL-1 或手术诱导的 OA 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中的 OPG 表达。 AnxA5 和 AnxA6 过度表达软骨细胞，并最终导致 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 病理学过程中刺激软骨破坏提供新的机制，并为 OA 的治疗提供新的治疗靶点。