Role of CCR2 in osteoarthritis

CCR2 在骨关节炎中的作用

• 批准号: 9751766
• 负责人: Lara Longobardi
• 金额: $ 34.21万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9751766
• 关键词: 3-Dimensional; Address; Affect; Age; Aging; Agonist; Ankle; Arthralgia; Arthritis; Binding; Biological Models; Bone Pain; Bone Spur; Bone remodeling; CC chemokine receptor 2; CCL2 gene; Cartilage; Cartilage injury; Cells; Chondrocytes; Clinic; Cohort Studies; Communities; Data; Degenerative polyarthritis; Development; Diagnostic radiologic examination; Early Diagnosis; Early Intervention; Early treatment; Environment; Evaluation; Event; Functional disorder; Gene Expression Profile; Genes; Genetic Recombination; Germ Lines; Goals; Hip region structure; Homeostasis; Homologous Gene; Human; In Vitro; Infiltration; Inflammation; Injections; Injury; Joints; Knee; Knee Osteoarthritis; Knock-out; Knowledge; Lead; Ligands; Link; Matrix Metalloproteinases; Measurable; Mechanics; Medial meniscus structure; Mediating; Mediator of activation protein; Medical; Modeling; Molecular; Mus; Myelogenous; Osteoblasts; Osteoclasts; Outcome; Pain; Pathogenesis; Pathologic; Pathway interactions; Patients; Pattern; Periosteum; Play; Pre-Clinical Model; Process; Production; Public Health; Replacement Arthroplasty; Reporting; Research; Role; Sclerosis; Serum; Severities; Signal Pathway; Signal Transduction; Structure; Synovitis; Testing; Time; Tissues; Trauma; Treatment Cost; Work; aggrecan; articular cartilage; base; behavioral response; bone; chemokine; clinical development; follow-up; histological studies; in vivo; inhibitor/antagonist; insight; joint destruction; joint injury; macrophage; mouse model; novel; organizational structure; osteoarthritis pain; pain behavior; pain inhibition; pain perception; pain reduction; pre-clinical; prevent; protective effect; receptor; response; young adult

PROJECT ABSTRACT Despite evidence that chemokines are increased in OA joints, few studies have examined their role in OA and the mechanism by which they promote joint destruction. The goal of this proposal is to determine if the CC- chemokine receptor 2 (CCR2) is a critical mediator of joint damage and pain in OA. Our preliminary studies and recent work by others suggest CCR2 inhibition may reduce both structural damage and pain in OA. If this is true, it would be a major breakthrough in the management of OA. However, more work is needed to confirm these findings and determine the optimal time to inhibit CCR2 during the OA process. Our data from a community- based cohort study of human OA shows that higher baseline serum levels of human CCL2 (which binds CCR2 receptor) correlates with progression of radiographic OA at 5-years follow up. Using the destabilized medial meniscus (DMM) murine model of injury-induced OA, we found that expression of CCL12 (the functional homologue of hCCL2), a CCR2 ligand we have previously found to be critical in joint development, increases in articular cartilage and bone during early OA. OA changes in articular cartilage (cartilage loss, increased MMP13 levels) and bone (bone sclerosis, osteophytes) and pain behavior were ameliorated by systemic blockade of the CCR2, if given for 4wks total within the first 8wks after DMM. However, if treatment was maintained for 8wks or more it was less effective on subsequent structural progression while still inhibiting pain, suggesting a disconnect between structure and pain that has been noted in human OA. These data suggest that CCR2 signaling has cellular and stage specific roles in OA, with beneficial effects when targeted at early stages. We hypothesize that injury-induced alterations in joint mechanics upregulate the CCR2 pathway in cartilage and bone in a time- dependent manner altering the structural organization of both tissues, ultimately leading to OA and pain. We propose to determine the role of dysregulated CCR2 signaling in cartilage at sequential times during injury- and age-induced OA, in order to establish the contribution of this tissue to the whole joint degeneration and OA pain (Aim 1). We will also evaluate whether osteoblast expression of CCR2 contributes to joint damage and pain during injury-induced OA (Aim 2). To accomplish this, we will obtain an inducible tissue-specific CCR2 deletion in chondrocytes (Aim 1) and osteoblasts (Aim 2) by combining chondrocyte- or osteoblast-specific expression of CreER with Tam injections in CCR2flox/flox mice crossed with Aggrecan-CreERT2 or Col1α1CreER, respectively. Because the myeloid infiltration consisting of macrophages and osteoclasts may cause accelerated OA bone remodeling that may reflect on bone damage and, indirectly, on cartilage homeostasis, we will also conditionally inactivate CCR2 in macropahges/osteoclasts (Aim 3) by crossing CCR2flox/flox mice with LysMERCre. We will also evaluate potential pathways altered by each tissue-selective CCR2 deletions. In addition, by using in-vitro human primary chondrocyte cultures, we will study the signaling pathways by which activation of CCR2 with its multiple ligands stimulates MMP production.

项目摘要 尽管有证据表明趋化因子在 OA 关节中增加，但很少有研究探讨它们在 OA 和 OA 中的作用。 他们促进联合破坏的机制。该提案的目标是确定 CC- 趋化因子受体 2 (CCR2) 是 OA 关节损伤和疼痛的关键介质。我们的初步研究和 其他人最近的研究表明，抑制 CCR2 可能会减少 OA 的结构损伤和疼痛。如果这是真的， 这将是OA管理的重大突破。然而，还需要更多的工作来证实这些 研究结果并确定 OA 过程中抑制 CCR2 的最佳时间。我们的数据来自社区 - 基于人类 OA 的队列研究表明，人类 CCL2（与 CCR2 结合）的基线血清水平较高 受体）与 5 年随访时放射学 OA 的进展相关。使用不稳定的内侧 在损伤诱导的 OA 半月板 (DMM) 小鼠模型中，我们发现 CCL12（功能性 hCCL2 的同源物）（我们之前发现的一种 CCR2 配体，对于关节发育至关重要）增加了 早期 OA 期间的关节软骨和骨。关节软骨的 OA 变化（软骨损失、MMP13 增加 水平）和骨骼（骨质硬化、骨赘）和疼痛行为通过系统性阻断而得到改善 CCR2，如果在 DMM 后的前 8 周内总共给予 4 周。然而，如果治疗持续8周或 更多的是，它对随后的结构进展的效果较差，同时仍然抑制疼痛，表明脱节 在人类 OA 中已经注意到结构和疼痛之间的关系。这些数据表明 CCR2 信号传导 OA 中的细胞和阶段特定作用，在早期阶段针对时可产生有益效果。我们假设 损伤引起的关节力学改变会在一段时间内上调软骨和骨中的 CCR2 通路 依赖的方式改变两个组织的结构组织，最终导致骨关节炎和疼痛。我们 提议确定在损伤和损伤期间连续时间软骨中失调的 CCR2 信号传导的作用 年龄诱发的 OA，以确定该组织对整个关节退化和 OA 疼痛的贡献 （目标 1）。我们还将评估 CCR2 的成骨细胞表达是否会导致关节损伤和疼痛 损伤引起的 OA 期间（目标 2）。为了实现这一目标，我们将获得可诱导的组织特异性 CCR2 缺失 通过结合软骨细胞或成骨细胞特异性表达，在软骨细胞（目标 1）和成骨细胞（目标 2）中 在分别与 Aggrecan-CreERT2 或 Col1α1CreER 杂交的 CCR2flox/flox 小鼠中注射 CreER 和 Tam。 因为巨噬细胞和破骨细胞组成的骨髓浸润可能会导致OA骨加速 可能反映骨损伤并间接影响软骨稳态的重塑，我们也会有条件地 通过将 CCR2flox/flox 小鼠与 LysMERCre 杂交，灭活巨噬细胞/破骨细胞中的 CCR2（目标 3）。我们将 还评估每个组织选择性 CCR2 缺失改变的潜在途径。此外，通过使用体外 人类原代软骨细胞培养物中，我们将研究 CCR2 及其激活的信号通路 多个配体刺激 MMP 的产生。