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) 吸收骨同时抑制骨质流失，从而导致骨质流失 成骨细胞 (OB) 构建骨骼的能力。重要的是，RA 患者会出现全身性的症状。 目前的治疗药物不能很好地控制骨质减少/骨质疏松症。因此，至关重要的是 开发在炎症性关节炎中促进骨骼合成代谢的新药物。此前，我们确定了 接头蛋白 Schnurri-3 (SHN3) 作为骨形成的有效抑制剂。缺乏SHN3的小鼠会产生 骨量逐渐增加，因为 SHN3 缺失增强了 OB 功能。有趣的是，RA 患者表示 SHN3 存在于发炎滑膜内和血清中的细胞中，并且 SHN3 表达在体外 OB 和 从 RA 患者中分离出的滑膜细胞 (FLS)，对 RA 相关细胞因子 TNF 和 IL-17A 产生反应。 值得注意的是，我们的初步数据表明 SHN3 缺乏可以防止炎症引起的骨质流失 体外和体内。因此，抑制SHN3是促进骨形成的一个有吸引力的机制。 治疗伴随 RA 的局部和全身骨质流失。最后，我们开发了一种针对骨骼的 重组腺相关病毒 (rAAV) 靶向 SHN3，可以预防或治疗 RA 中的骨质流失。 目标 1 将测试 SHN3 缺陷是否可以限制 TNF-α 中骨质疏松症和骨侵蚀的发展。 通过增强 OB 功能诱导 RA 模型，并且可以在血清转移中促进关节糜烂的愈合 诱发关节炎模型。此外，我们将检验以下假设：OB 中 SHN3 缺陷会阻止 RA 相关细胞因子 TNF 和 IL-17A 抑制成骨，而 FLS 缺乏 抑制炎症细胞因子和 WNT 拮抗剂的产生，从而抑制 OB 差异化。目标 2 将确定 SHN3 缺陷预防的分子机制 炎症诱导的成骨抑制，以确定促进骨形成的新靶点。我们将 检查 RA 相关细胞因子 TNF 和 IL-17A 如何通过 NF-诱导 SHN3 转录物上调 kB 途径和通过 ERK MAPK 途径磷酸化 SHN3 的稳定性，从而抑制 WNT/b-连环蛋白途径和 OB 分化。目标 3 将确定骨特异性 rAAV 是否​​介导 体内 SHN3 沉默可预防 RA 中炎症引起的骨质流失。全身性骨质流失和关节骨质流失 将在用携带 SHN3 沉默子的骨特异性 rAAV 治疗的 RA 小鼠模型中对侵蚀进行量化。 转录组分析将在从 RA 小鼠模型中分离的 AAV 转导的 OB 谱系细胞中进行 鉴定 SHN3 通路中促进炎症诱导骨愈合的潜在分子效应器 损失。这项工作的成功完成将为 SHN3 缺陷可以提供原理证明 增强 RA 炎症引起的骨质流失部位的骨形成。了解 SHN3 的原理 抑制可防止在这种情况下抑制 OB 分化并识别潜在的新 SHN3 通路中的调节因子应该为炎症性关节炎提供重要的治疗靶点。