Regulatory Mechanisms of Implant-Induced Osteolysis

植入物引起的骨溶解的调节机制

• 批准号: 8294459
• 负责人: YOUSEF ABU-AMER
• 金额: $ 32.83万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8294459
• 关键词: Abbreviations; Address; Aging; Arthritis; Attenuated; Binding; Binding Proteins; Bone Marrow; Calvaria; Cells; Complex; Degenerative Disorder; Deubiquitination; Diagnosis; Event; Failure; Hemagglutinin; Implant; In Vitro; Inflammatory; Inflammatory Response; Interleukin-1; Investigation; Joints; Lead; Ligase; Link; Lysine; MAP Kinase Gene; MAP kinase kinase kinase 7; MAP3K7 gene; MAPK8 gene; Mediating; Mediator of activation protein; Mitogen-Activated Protein Kinases; Mitogens; Molecular; Morbidity - disease rate; Mus; Mutant Strains Mice; NF-kappa B; Operative Surgical Procedures; Orthopedics; Osteoclasts; Osteolysis; Osteolytic; Osteoporosis; Pathologic; Pathway interactions; Patients; Peptides; Phosphotransferases; Polymethyl Methacrylate; Polyubiquitination; Preventive; Process; Protein Kinase; Proteins; Regulation; Replacement Arthroplasty; Rheumatoid Arthritis; Role; Signal Transduction; Small Interfering RNA; TNF Receptor-Associated Factors; TNF gene; TNFSF11 gene; TRAF6 gene; Testing; Therapeutic; Time; Transcription Factor AP-1; Ubiquitin; Ubiquitin-Conjugating Enzymes; Ubiquitination; Weight-Bearing state; Work; base; bone; human RIPK1 protein; in vivo; inhibitor/antagonist; macrophage; mortality; mouse model; mutant; novel; osteoclastogenesis; particle; prevent; public health relevance; receptor; response; stress-activated protein kinase 1; tartrate-resistant acid phosphatase; ubiquitin ligase

DESCRIPTION (provided by applicant): Degenerative conditions of large weight bearing joints resulting from aging and/or pathologic conditions such as arthritis and osteoporosis lead to surgical intervention approaches chiefly total joint replacement (TJR). However, implant-derived wear debris occurs with time causing inflammatory responses culminating with osteolysis and failure of implants. Subsequent revision surgery of the failing joint implant, is often more difficult, and associated with increased morbidity and mortality especially for aging patients with weaker bones. Therefore, the need for effective approaches to diagnose, prevent, and/or treat complications of TJR have risen in recent years. Thus, better understanding of the processes and mechanisms underlying pathologic and osteolytic events leading to joint failure is essential to provide appropriate preventive and therapeutic countermeasures. The pathologic response to implant wear-debris constitutes a major component of this phenomenon and is under intense investigation. Recent work by several groups including ours has identified important cellular entities and secreted factors that contribute to inflammatory osteolysis. In previous work, we have shown that PMMA particles contribute to inflammatory osteolysis through stimulation of major pathways in osteoclast precursors, primarily NF-?B and MAP kinases. The former pathway requires assembly of large IKK complex encompassing IKK1, IKK2, and IKK?, also known as NEMO. We have shown recently that interfering with the NF-?B and MAPK activation pathways, through introduction of inhibitors and decoy molecules, impede PMMA-induced osteolysis in mouse models of experimental calvarial osteolysis and inflammatory arthritis. In our recent work, we found that PMMA particles directly activate the upstream transforming growth factor beta activated kinase-1 (TAK1) which is a key regulator of signal transduction cascades leading to activation of NF-?B and AP-1 factors. More importantly, we found that PMMA particles induce TAK1 binding to NEMO, RIP1, and UBC13. In addition, we show that PMMA particles induced TRAF6 binding to NEMO and lack of TRAF6 significantly attenuates NEMO ubiquitination. We further demonstrate that PMMA induction of NF-?B and MAPK is impaired in TAK1-null and NEMO mutant cells. These responses were not aided by TNF or RANKL. Altogether, these results led us to hypothesize that PMMA particles maybe inducing K63-linked ubiquitination of NEMO, RIP1, and other target proteins, events likely mediated by TRAF6, TAK1 and UBC13. Relevant to this hypothesis, it has been documented that a key mediator of LPS, IL-1, and RANKL signaling, namely TRAF6, is ubiquitin ligase. In separate studies, it was further established that a variety of upstream signals augment ubiquitination-based signaling network dominated by TAK1/TABs/RIP/NEMO/UBC complex. Thus, we propose to investigate the following specific aims: 1. Delineate the molecular steps underlying PMMA-induced regulation of NEMO. 2. Investigate if TRAF6/TAK1/NEMO POLYUBIQUITINATION events mediate PMMA-induced osteoclastogenesis. 3. Determine the effect of inhibiting POLY-UB-NEMO signaling on PMMA-induced calvarial osteolysis. PUBLIC HEALTH RELEVANCE: Total joint implant failure is attributed, at least in part, to orthopedic particle-induced osteolysis. The mechanisms underlying this pathologic condition remain unclear. Our recent work has implicated heightened osteoclast activity, owing to enhanced intracellular activation of NF-?B and MAP kinase pathways, as the leading cause for inflammatory osteolysis. We provide evidence that key mediators of these two pathways, including TGF2-activating kinase (TAK1), IKK?/NEMO, E2-ligases such as Ubc13, and key ubiquitin-dependent events are induced by polymethylmethacrylate (PMMA) particles in osteoclast precursors. Thus, we will utilize in vitro and in-vivo approaches including genetically-modified mice to examine the role of ubiquitin-mediated events that contribute to exacerbation of PMMA-induced inflammatory osteolysis. Our proposal addresses novel questions and holds promise to identify novel selective anti-osteolytic therapies.

描述（由申请人提供）：由衰老和/或病理状况（例如关节炎和骨质疏松症）引起的大承重关节的退化条件，导致手术干预方法主要是总关节置换（TJR）。然而，随着时间的流逝，植入物衍生的磨损碎屑发生，导致炎症反应最终导致溶解和植入物的失败。随后的失败关节植入物的修订手术通常更加困难，并且与发病率和死亡率升高有关，尤其是对于骨骼较弱的衰老患者而言。因此，近年来需要有效的诊断，预防和/或治疗并发症的方法的需求。因此，更好地理解导致关节失败的病理和骨化事件的过程和机制对于提供适当的预防性和治疗性对策至关重要。对植入物磨损的病理反应构成了这种现象的主要组成部分，并且正在接受激烈的研究。包括我们的几个小组的最新工作确定了有助于炎症骨溶解的重要细胞实体和分泌因素。在先前的工作中，我们已经表明，PMMA颗粒通过刺激破骨细胞前体的主要途径（主要是NF-？b和MAP激酶）导致炎症性溶解。以前的途径需要组装大型IKK复合物，包括IKK1，IKK2和IKK？，也称为Nemo。我们最近表明，通过引入抑制剂和诱饵分子来干扰NF-？B和MAPK激活途径，在实验性钙钙骨溶解和炎症性关节炎的实验性小鼠模型中阻碍了PMMA诱导的骨溶解。在我们最近的工作中，我们发现PMMA颗粒直接激活上游转化生长因子β激活的激酶-1（TAK1），这是信号转导级联反应的关键调节剂，导致NF-？B和AP-1因子激活。更重要的是，我们发现PMMA颗粒诱导TAK1与Nemo，RIP1和UBC13结合。此外，我们表明PMMA颗粒诱导TRAF6与Nemo的结合，而缺乏TRAF6会显着减弱Nemo的泛素化。我们进一步证明了Nf- b的PMMA诱导和MAPK在TAK1-NULL和NEMO突变细胞中受到损害。 TNF或RANKL没有帮助这些响应。总的来说，这些结果使我们假设PMMA颗粒可能诱导NeMo，RIP1和其他靶蛋白的K63连接的泛素化，这可能是由TRAF6，TAK1和UBC13介导的事件。与该假设相关的是，已经证明了LPS，IL-1和RANKL信号的关键介体，即TRAF6是泛素连接酶。在单独的研究中，进一步确定了各种以TAK1/TABS/RIP/RIP/NEMO/UBC COMPLECT为主导的基于泛素化的信号网络增强基于泛素化的信号网络。因此，我们建议研究以下特定目的：1。描述PMMA诱导的NEMO调控的基础分子步骤。 2。研究TRAF6/TAK1/NEMO多泛素化事件是否介导PMMA诱导的破骨细胞生成。 3。确定抑制聚-UB-NEMO信号对PMMA诱导的颅骨骨溶解的影响。 公共卫生相关性：至少部分归因于骨科诱导的骨溶解的总植入物衰竭。这种病理状况的基础机制尚不清楚。由于NF-？B和MAP激酶途径的细胞内激活增强，我们最近的工作暗示了破骨细胞活性的增强，这是炎症骨溶解的主要原因。我们提供的证据表明，这两种途径的关键介体，包括TGF2激活激酶（TAK1），IKK？/NEMO，E2-ligases，例如UBC13，以及关键的泛素依赖性事件，是由多甲基甲基丙烯酸酯（PMMA）颗粒（PMMA）颗粒诱导的。因此，我们将利用包括遗传改性小鼠在内的体外和体内方法来检查泛素介导的事件的作用，这有助于加剧PMMA诱导的炎性骨溶解。我们的建议解决了新的问题，并有望确定新颖的选择性抗固定疗法。