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: 10052718
• 负责人: Carla Rose Scanzello
• 金额: $ 49.39万
• 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/10052718
• 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; Effectiveness; 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; 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，导致慢性进行性关节疼痛和组织 整个关节损坏。目前没有医疗治疗可有效防止进行性关节 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疼痛和病理学的主要驱动因素，目前尚不清楚哪种分子途径直接 驱动慢性OA疼痛和疾病进展，或者是治疗发育的关键靶标。我们的实验室有 表明CD14的不足是巨噬细胞和 其他髓样细胞类型，可以防止与OA相关的骨骼复制和与疼痛相关的关节功能障碍 小鼠膝盖受伤后。在患者中，关节液中的CD14增加并与关节内有关 巨噬细胞浸润以及疼痛的严重程度。该受体促进了Toll样受体（TLR）信号传导。 TLR是先天免疫传感器，这是慢性炎症的重要初始触发因素 非感染组织损伤。此外，我们的团队最近证明了TLR信号对 通过直接刺激疼痛传递（伤害感受）神经元的膝关节疼痛的发育 支撑膝盖的背根神经节（DRG）。在此提案中，我们将测试CD14上的假设 髓样细胞在内 炎症），而CD14也通过直接使神经元支配神经来增加OA疼痛 关节炎关节。我们将利用体外技术和内侧半月板的鼠不稳定（DMM） OA的模型，了解CD14驱动OA病理学和的细胞和分子机制 关节组织中的炎症。具体而言，在AIM 1中，我们将使用多学科的方法来确定如何 CD14的遗传缺乏可修饰OA进展过程中的炎症和骨骼复制，并使用 DMM型号。我们将确定髓样细胞对在炎症和骨造成的贡献 使用骨髓（BM）嵌合小鼠进行模型。最后，我们将测试CD14对分化的影响 驱动骨骼复制的细胞（破骨细胞）和髓样疼痛炎症介质的产生 细胞类型。在AIM 2中，我们将表征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响应，比较WT和WT 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，并将其进行比较 预计DRG反应和疼痛都会在CD14缺乏菌株中钝化。最后，在目标3中，我们将 测试CD14的药理靶向是否会减少DMM诱导损伤后的OA进展和疼痛。 然后，这项研究将指定分子和细胞框架以设计未来的抗炎 OA针对CD14和TLR介导的机制的治疗剂。