Novel approaches to promote healing of bone loss in inflammatory arthritis

促进炎症性关节炎骨质流失愈合的新方法

• 批准号: 10590694
• 负责人: Ellen M Gravallese
• 金额: $ 52.44万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-14 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10590694
• 关键词: Ablation; Adaptor Signaling Protein; Alleles; Anabolic Agents; Antibodies; Arthritis; Binding; Binding Sites; Biochemical; Biological; Bone Marrow; Bone Resorption; Bone necrosis; Cell Culture Techniques; Cell Lineage; Cell Separation; Cells; Clinical; Data; Development; Disease; Fracture; Genetic Transcription; Grant; Histologic; Histology; Human; IL17 gene; In Vitro; Inflammation; Inflammatory; Inflammatory Arthritis; Jaw; Joints; Knee joint; Knock-in; Link; MAP Kinase Gene; Mass Spectrum Analysis; Mediating; Messenger RNA; Modeling; Molecular; Mus; Mutation; NF-kappa B; New Agents; Osteoblasts; Osteoclasts; Osteogenesis; Osteopenia; Osteoporosis; Osteoporosis prevention; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Phosphorylation; Production; Proteins; RNA Interference; Recombinant adeno-associated virus (rAAV); Resolution; Rheumatoid Arthritis; Serum; Site; Sorting; Stromal Cells; Synovitis; TNF gene; Testing; Therapeutic Agents; Therapeutic Uses; Transcript; Up-Regulation; Validation; WNT Signaling Pathway; Work; adeno-associated viral vector; ankle joint; antagonist; beta catenin; bone; bone erosion; bone healing; bone loss; bone mass; bone repair; cytokine; experimental study; gene therapy; healing; in vitro Model; in vivo; inhibitor; innovation; joint inflammation; long bone; microCT; mouse model; mutant; novel; novel strategies; novel therapeutics; osteoblast differentiation; overexpression; prevent; promoter; response; side effect; spine bone structure; therapeutic target; transcriptomic profiling

Rheumatoid arthritis (RA) leads to bone loss by activating osteoclasts (OCs) to resorb bone while suppressing the ability of osteoblasts (OBs) to build bone. Importantly, patients with RA develop systemic osteopenia/osteoporosis that is not well controlled by current therapeutic agents. Therefore, it is critical to develop new agents that are anabolic for bone in the setting of inflammatory arthritis. Previously, we identified the adaptor protein Schnurri-3 (SHN3) as a potent inhibitor of bone formation. Mice lacking SHN3 develop a progressive increase in bone mass, as SHN3 deletion enhances OB function. Intriguingly, RA patients express SHN3 in cells within inflamed synovium and in serum, and SHN3 expression is induced in vitro in OBs and in synoviocytes (FLS) isolated from RA patients in response to the RA-associated cytokines TNF and IL-17A. Notably, our preliminary data show that SHN3-deficiency can protect from inflammation-induced bone loss in vitro and in vivo. Therefore, inhibition of SHN3 is an attractive mechanism to promote bone formation to treat the local and systemic bone loss that accompanies RA. Finally, we have developed a bone-specific recombinant adeno-associated virus (rAAV) that targets SHN3 and could prevent or treat bone loss in RA. Aim 1 will test whether SHN3-deficiency can limit the development of osteoporosis and bone erosion in a TNF- induced RA model by augmenting OB function, and can promote healing of articular erosions in a serum transfer- induced arthritis model. Additionally, we will test the hypothesis that SHN3-deficiency in OBs prevents the suppression of osteogenesis by the RA-associated cytokines TNF and IL-17A, while its deficiency in FLS suppresses production of inflammatory cytokines and WNT antagonists that lead to inhibition of OB differentiation. Aim 2 will determine the molecular mechanisms by which SHN3 deficiency protects from inflammation-induced suppression of osteogenesis to identify novel targets promoting bone formation. We will examine how the RA-associated cytokines TNF and IL-17A induce upregulation of SHN3 transcripts via the NF- kB pathway and stabilization of phosphorylated SHN3 via the ERK MAPK pathway, resulting in suppression of the WNT/b-catenin pathway and OB differentiation. Aim 3 will determine whether bone-specific rAAV-mediated silencing of SHN3 in vivo prevents inflammation-induced bone loss in RA. Systemic bone loss and articular erosion will be quantified in mouse models of RA treated with the bone-specific rAAV carrying a SHN3 silencer. Transcriptomic profiling will be performed in AAV-transduced OB-lineage cells isolated from RA mouse models to identify potential molecular effectors in the SHN3 pathway that promote healing of inflammation-induced bone loss. Successful completion of this work will provide proof-of-principle that SHN3-deficiency can augment bone formation at sites of inflammation-induced bone loss in RA. Understanding how SHN3 inhibition protects from suppression of OB differentiation in this setting and identifying potential novel regulators in the SHN3 pathway should provide important therapeutic targets for inflammatory arthritis.

类风湿性关节炎(RA)通过激活破骨细胞(OCs)吸收骨而导致骨丢失 成骨细胞(OBS)建骨的能力。重要的是，类风湿关节炎患者发展为全身性 骨量减少/骨质疏松症，目前的治疗药物不能很好地控制它。因此，至关重要的是 在炎症性关节炎的背景下，开发对骨骼合成代谢的新试剂。此前，我们确定了 适配蛋白SchNurri-3(SHN3)是一种有效的骨形成抑制因子。缺乏SHN3的小鼠会患上 骨量进行性增加，因为SHN3缺失增强了OB功能。耐人寻味的是，RA患者表达 SHN3在炎症滑膜细胞和血清中表达，在OBS和OBS中体外诱导SHN3表达 从RA患者分离的滑膜细胞(FLS)对RA相关细胞因子TNF和IL-17A的反应。 值得注意的是，我们的初步数据显示，SHN3缺乏可以预防炎症诱导的骨丢失 体外和体内。因此，抑制SHN3是促进骨形成的一种诱人机制。 治疗伴随类风湿关节炎的局部和全身骨丢失。最后，我们开发了一种针对骨骼的 针对SHN3的重组腺相关病毒(RAAV)，可预防或治疗RA的骨丢失。 目的1将测试SHN3缺乏是否可以限制骨质疏松症和骨侵蚀的发展，在一个肿瘤坏死因子- 通过增强OB功能诱导RA模型，并能促进血清移植中关节糜烂的愈合 诱导关节炎模型。此外，我们还将检验这一假设，即OBS中SHN3缺乏会阻止 类风湿关节炎相关细胞因子TNF和IL-17A对成骨的抑制作用及其在FLS中的缺陷 抑制炎性细胞因子和WNT拮抗剂的产生，从而抑制OB 差异化。目标2将确定SHN3缺乏保护的分子机制 炎症诱导的成骨抑制以确定促进骨形成的新靶点。我们会 研究RA相关细胞因子肿瘤坏死因子和白介素17A如何通过核因子-1诱导SHN3转录上调 Kb途径和通过ERK MAPK途径稳定磷酸化的SHN3，导致抑制 Wnt/b-catenin途径与OB分化AIM 3将确定骨特异性rAAV介导的 体内沉默SHN3可预防类风湿性关节炎引起的骨丢失。全身性骨丢失与关节 在使用携带SHN3消音器的骨骼特异性rAAV治疗的RA小鼠模型中，侵蚀将被量化。 将在从RA小鼠模型分离的AAV转导的OB系细胞中进行转录图谱分析 确定SHN3通路中促进炎症诱导骨愈合的潜在分子效应物 损失。这项工作的成功完成将提供SHN3缺陷可以 促进炎症引起的RA骨丢失部位的骨形成。了解SHN3如何 抑制在这种背景下保护OB分化不受抑制，并识别潜在的小说 SHN3途径的调节因子应该为炎症性关节炎提供重要的治疗靶点。