Modulation of Inflammation in Osteoarthritis via CD14-mediated pattern recognition

通过 CD14 介导的模式识别调节骨关节炎炎症

• 批准号: 10224102
• 负责人: Carla Rose Scanzello
• 金额: $ 45.37万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10224102
• 关键词: Adult; Analgesics; Anti-Inflammatory Agents; Arthralgia; Arthritis; Behavior; Biology; Blocking Antibodies; Bone Marrow; Bone remodeling; CD14 Antigen; CD14 gene; Cartilage; Cell Differentiation process; Cells; Characteristics; Chimera organism; Chronic; Coupled; Cytometry; Degenerative polyarthritis; Dependence; Deterioration; Development; Disease; Disease Progression; Fatty acid glycerol esters; Functional disorder; Future; Genetic; Hematopoietic; Hematopoietic System; Hyperalgesia; Image; Immune; Immunology; In Vitro; Infection; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Injections; Injury; Joints; Knee; Knee Injuries; Knee Osteoarthritis; Laboratories; Lead; Leukocytes; Ligands; Liquid substance; Measures; Medial meniscus structure; Mediating; Mediator of activation protein; Medical; Modeling; Molecular; Mus; Myelogenous; Myeloid Cells; Neurons; Nociception; Opioid; Osteitis; Osteoclasts; Outcome; Pain; Pathologic; Pathology; Pathway interactions; Patients; Pattern Recognition; Pattern recognition receptor; Pharmaceutical Preparations; Pharmacology; Positioning Attribute; Production; Quality of life; Reagent; Receptor Signaling; Regulation; Rehabilitation therapy; Replacement Arthroplasty; Research Personnel; Role; Severities; Signal Transduction; Specific qualifier value; Spinal Ganglia; Stimulus; Symptoms; Synovial Membrane; Techniques; Testing; Therapeutic; Tissues; Toll-like receptors; Treatment Efficacy; Treatment Protocols; Work; arthropathies; bone; cell type; cellular targeting; clinical translation; cost; design; disability; effectiveness evaluation; experimental study; improved; in vivo; inflammatory pain; interdisciplinary approach; joint destruction; joint injury; macrophage; monocyte; mouse model; mutant; new therapeutic target; novel strategies; osteoarthritis pain; pain behavior; pain reduction; pressure; prevent; receptor; receptor-mediated signaling; release of sequestered calcium ion into cytoplasm; response; sensor; side effect; targeted treatment; therapeutic development; therapeutic evaluation; tissue injury

ABSTRACT Osteoarthritis (OA) is a leading cause of disability in adults, causing chronic progressive joint pain and tissue damage throughout the joint. No current medical treatments are effective at preventing the progressive joint deterioration, pain and disability characteristic of OA. Although it is known that inflammation and bone- remodeling are major drivers of OA pain and pathology, it is not yet clear which molecular pathways directly drive chronic OA pain and disease progression or are key targets for therapeutic development. Our lab has shown that deficiency of CD14, an inflammatory pattern-recognition receptor expressed by macrophages and other myeloid cell types, protects against OA-associated bone-remodeling and pain-related joint dysfunction after knee injury in mice. In patients, CD14 is increased in joint fluid and associated with intra-articular macrophage infiltration as well as pain severity. This receptor facilitates Toll-like receptor (TLR) signaling. TLRs are innate immune sensors which are important initial triggers of chronic inflammation in response to non-infectious tissue damage. In addition, our team has recently demonstrated that TLR-signaling is critical to development of knee OA pain in mice, via direct stimulation of pain-transmitting (nociceptive) neurons in the dorsal root ganglia (DRG) that innervate the knee. In this proposal, we will test the hypothesis that CD14 on myeloid cells including macrophages promotes OA pain-related pathology (bone remodelling and inflammation), while CD14 also augments OA pain by directly sensitizing neurons innervating the arthritic joint. We will utilize in vitro techniques and the murine destabilization of the medial meniscus (DMM) model of OA, to understand the cellular and molecular mechanisms by which CD14 drives OA pathology and inflammation in joint tissues. Specifically, in Aim 1 we will use a multi-disciplinary approach to determine how genetic deficiency of CD14 modifies inflammation and bone-remodeling during progression of OA, using the DMM model. We will determine the contribution of myeloid cells to inflammation and bone-remodeling in the model by using bone marrow (BM) chimeric mice. Lastly, we will test the effects of CD14 on differentiation of cells that drive bone-remodeling (osteoclasts) and production of inflammatory mediators of pain from myeloid cell types. In Aim 2 we will characterize effects of CD14 on TLR-mediated DRG neuronal activation and joint pain. We will use TLR stimuli with relevance to OA to evaluate DRG responses in vitro, comparing WT and CD14-deficient cells. We will additionally compare pain responses to injection of TLR-stimuli into the joint, and expect that both DRG responses and pain will be blunted in the CD14 deficient strain. Finally, in Aim 3 we will test whether pharmacological targeting of CD14 reduces OA progression and pain after DMM-induced injury. This study will then specify the molecular and cellular framework to design future anti-inflammatory therapeutics for OA aimed at CD14- and TLR-mediated mechanisms.

抽象的 骨关节炎 (OA) 是成人残疾的主要原因，会导致慢性进行性关节疼痛和组织损伤 整个关节的损伤。目前没有任何药物治疗可以有效预防进行性关节 OA 的恶化、疼痛和残疾特征。尽管众所周知，炎症和骨 重塑是 OA 疼痛和病理的主要驱动因素，目前尚不清楚哪些分子途径直接影响 驱动慢性 OA 疼痛和疾病进展，或者是治疗开发的关键目标。我们实验室有 研究表明，CD14（一种由巨噬细胞表达的炎症模式识别受体）的缺陷 其他骨髓细胞类型，防止骨关节炎相关的骨重塑和疼痛相关的关节功能障碍 小鼠膝盖受伤后。在患者中，关节液中 CD14 增加，并与关节内 巨噬细胞浸润以及疼痛严重程度。该受体促进 Toll 样受体 (TLR) 信号传导。 TLR 是先天免疫传感器，是响应慢性炎症的重要初始触发因素。 非感染性组织损伤。此外，我们的团队最近证明 TLR 信号传导对于 通过直接刺激小鼠膝关节中的疼痛传递（伤害感受）神经元来引起小鼠膝关节 OA 疼痛 支配膝盖的背根神经节（DRG）。在这个提案中，我们将测试 CD14 的假设 包括巨噬细胞在内的骨髓细胞促进 OA 疼痛相关病理（骨重塑和 炎症），而 CD14 还可以通过直接使支配 OA 的神经元变得敏感来增强 OA 疼痛。 关节炎关节。我们将利用体外技术和小鼠内侧半月板不稳定 (DMM) OA 模型，了解 CD14 驱动 OA 病理的细胞和分子机制， 关节组织炎症。具体来说，在目标 1 中，我们将使用多学科方法来确定如何 CD14 的遗传缺陷改变了 OA 进展过程中的炎症和骨重塑，利用 数字万用表型号。我们将确定骨髓细胞对炎症和骨重塑的贡献 使用骨髓（BM）嵌合小鼠建立模型。最后，我们将测试CD14对分化的影响 驱动骨重塑的细胞（破骨细胞）和髓系疼痛炎症介质的产生 细胞类型。在目标 2 中，我们将描述 CD14 对 TLR 介导的 DRG 神经元激活和关节的影响 疼痛。我们将使用与 OA 相关的 TLR 刺激来评估体外 DRG 反应，比较 WT 和 CD14 缺陷细胞。我们还将比较将 TLR 刺激注射到关节中的疼痛反应，以及 预计 CD14 缺陷株的 DRG 反应和疼痛都会减弱。最后，在目标 3 中，我们将 测试 CD14 的药理靶向是否可以减少 DMM 引起的损伤后 OA 的进展和疼痛。 然后，这项研究将指定分子和细胞框架来设计未来的抗炎药物 针对 CD14 和 TLR 介导机制的 OA 治疗。