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: 10365023
• 负责人: Ellen M Gravallese
• 金额: $ 53.97万
• 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/10365023
• 关键词: Adaptor Signaling Protein; Alleles; Anabolic Agents; Antibodies; Arthritis; Binding; Binding Sites; Biochemical; Biological; Bone Marrow; Bone necrosis; Cell Culture Techniques; Cell Lineage; Cells; Clinical; Data; Development; Disease; Fracture; Genetic Transcription; Grant; Histologic; Histology; Human; In Vitro; Inflammation; Inflammatory; Inflammatory Arthritis; Interleukin-17; Jaw; Joints; Knee joint; Knock-in; Lead; 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; 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; base; 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; transcriptome; transcriptomics

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（OC）吸收骨，同时抑制 成骨细胞（OB）构建骨骼的能力。重要的是，RA患者会出现全身性疾病 目前的治疗剂不能很好地控制骨质减少/骨质疏松症。因此，关键是 在炎症性关节炎的情况下开发新的骨合成代谢剂。此前，我们发现 衔接蛋白Schnurri-3（SHN 3）作为骨形成的有效抑制剂。缺乏SHN 3的小鼠发育为 随着SHN 3缺失增强OB功能，骨量逐渐增加。有趣的是，RA患者表示 SHN 3在炎症滑膜和血清中的细胞中表达，并且SHN 3表达在体外OB和 从RA患者分离的滑膜细胞（FLS）对RA相关细胞因子TNF和IL-17 A的应答。 值得注意的是，我们的初步数据表明，SHN 3缺乏可以保护炎症诱导的骨丢失， 体外和体内。因此，抑制SHN 3是促进骨形成的有吸引力的机制， 治疗伴随RA的局部和全身骨质流失。最后，我们开发了一种骨特异性 重组腺相关病毒（rAAV）靶向SHN 3，可以预防或治疗RA的骨丢失。 目的1将测试SHN 3缺乏是否可以限制TNF-α诱导的骨质疏松症和骨侵蚀的发展。 通过增强OB功能诱导RA模型，并能促进血清转移中关节糜烂的愈合， 诱导性关节炎模型。此外，我们还将检验OBs中SHN 3缺陷会阻止 RA相关细胞因子TNF和IL-17 A抑制骨生成，而其在FLS中的缺陷 抑制导致OB抑制的炎性细胞因子和WNT拮抗剂的产生 分化目的2将确定SHN 3缺陷保护的分子机制， 炎症诱导的骨生成抑制，以鉴定促进骨形成的新靶点。我们将 研究RA相关细胞因子TNF和IL-17 A如何通过NF-κ B诱导SHN 3转录物的上调。 kB途径和通过ERK MAPK途径稳定磷酸化SHN 3，导致抑制 WNT/β-catenin通路与OB分化。目的3将确定骨特异性rAAV介导的 SHN 3在体内的沉默防止了RA中炎症诱导的骨丢失。全身性骨丢失和关节 将在用携带SHN 3沉默物的骨特异性rAAV处理的RA小鼠模型中定量侵蚀。 将在从RA小鼠模型分离的AAV转导的OB谱系细胞中进行转录组学分析。 鉴定SHN 3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SHN 3缺陷可以 增加RA炎症诱导骨丢失部位的骨形成。了解SHN 3 在这种情况下，抑制可防止OB分化受到抑制，并鉴定潜在的新的 SHN 3通路中的调节剂应该为炎性关节炎提供重要的治疗靶点。