Targeting hexokinase-2 in rheumatoid arthritis

靶向己糖激酶 2 治疗类风湿性关节炎

• 批准号: 10405768
• 负责人: Monica Guma
• 金额: $ 9.24万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10405768
• 关键词: Arthritis; Autoimmune Diseases; Binding; Biological Response Modifier Therapy; Biology; Cartilage; Cells; Combined Modality Therapy; Complement; Data; Development; Disease; Enzymes; Fibroblasts; Glycolysis Inhibition; Grant; Hexokinase 2; Homeostasis; Immunosuppression; Immunotherapy; Impairment; Inflammation; Interleukin-6; Intra-Articular Injections; Joints; Knee; Metabolic; Metabolic Pathway; Metabolism; Miconazole; Mitochondria; Mus; Normal Cell; Oncology; Pathogenesis; Pathway interactions; Patients; Peptides; Pharmaceutical Preparations; Phenotype; Protein Isoforms; Research; Rheumatoid Arthritis; Risk; Sampling; Severities; Synovial Cell; Synovial Membrane; Testing; Therapeutic; Thick; Tissues; Translating; Up-Regulation; adenoviral-mediated; arthropathies; bone; cell motility; cell type; extracellular; glucose metabolism; improved; insight; joint destruction; knock-down; macrophage; migration; mutant; novel; novel strategies; novel therapeutics; overexpression; small molecule

PROJECT SUMMARY After several years of research on biological therapies and small molecules to target inflammation, we need a different strategy to further get more insights into mechanisms underlying RA pathogenesis and identify potential new treatments, as a significant proportion of patients are partial responders. In other fields such as oncology the concept of metabolic reprograming to improve immunotherapy are concepts we truly believe should be translated into autoimmune diseases to complement current therapies. However, there are little data about targeting metabolic changes in RA. We seek with this grant a better understanding of the biology of these metabolic pathways in RA to better characterize a new approach in its therapeutic armamentarium. Our finding that hexokinase 2 (HK2) activity is enhanced only in RA synovium and in different RA synovial cells, suggest a cooperative metabolic reprograming in the joint that contributes to RA development and progression. Our preliminary data demonstrate that while HK1 expression is expressed in both OA and RA synovium, HK2 expression co-localizes with MO and FLS markers, and is only observed in RA and not in OA synovial samples. We also show that HK2 regulates key FLS function as HK2 knockdown impaired FLS invasion. Conversely, HK2 overexpression increases FLS invasion and migration rate. Of note, lactate and PLOD2, which are involved in cell migration and invasion, are upregulated after HK2 expression. Up-regulation of extracellular lactate also suggests a metabolic shift towards accelerated glycolytic metabolism. An HK2 mutant lacking its mitochondrial- binding motif (HK2ΔN) reversed the invasive phenotype. In MO, a peptide that dissociates HK2 from mitochondria, impaired IL-6 secretion. Importantly, adenovirus-mediated expression of HK2 in the knee by intra- articular injection induced synovial thickness, which was much less evident when HK2ΔN was intra-articular injected. Finally, HK2F/F-Col1a1 mice, which deletes HK2 in FLS among other non-hematopoietic cells, and treatment with clotrimazole, which dissociated HK2 from mitochondria, significantly decreased arthritis severity. Thus, we will test the hypothesis that mitochondrial HK2 is key regulator of FLS phenotype and MO activation, which contributes to joint destruction in RA. The identification of HK2, an isoform-specific contributor to elevated cell glucose metabolism in RA synovial tissue offers a safer approach than global glycolysis inhibition. HK2 could be selectively targeted without compromising systemic homeostasis or corresponding metabolic function in normal cells as a novel additional approach for combination therapy in RA joint disease independent of systemic immunosuppression.

项目概要 经过数年针对炎症的生物疗法和小分子的研究，我们需要一种 不同的策略，以进一步深入了解 RA 发病机制并确定潜在的 新的治疗方法，因为很大一部分患者是部分缓解者。在肿瘤学等其他领域 我们真正相信通过代谢重编程来改善免疫治疗的概念应该是 转化为自身免疫性疾病以补充当前的疗法。但有关资料却很少 针对 RA 的代谢变化。我们寻求通过这笔资助更好地了解这些生物的生物学 RA 的代谢途径，以更好地表征其治疗设备中的新方法。我们的发现 己糖激酶 2 (HK2) 活性仅在 RA 滑膜和不同 RA 滑膜细胞中增强，表明 关节内的协同代谢重编程有助于 RA 的发生和进展。我们的 初步数据表明，虽然 HK1 表达在 OA 和 RA 滑膜中表达，但 HK2 表达与 MO 和 FLS 标记共定位，并且仅在 RA 中观察到，而在 OA 滑膜样本中观察不到。 我们还表明，当 HK2 敲低损害 FLS 入侵时，HK2 调节关键的 FLS 功能。相反，HK2 过度表达会增加 FLS 侵袭和迁移率。值得注意的是，乳酸和 PLOD2 参与 HK2表达后细胞迁移和侵袭上调。细胞外乳酸也上调 表明代谢转向加速糖酵解代谢。缺乏线粒体的 HK2 突变体 结合基序（HK2ΔN）逆转了侵袭表型。在 MO 中，一种将 HK2 从 线粒体，IL-6 分泌受损。重要的是，腺病毒介导的 HK2 在膝关节内的表达 关节注射引起滑膜厚度，当 HK2ΔN 位于关节内时，这种情况不太明显 注射。最后，HK2F/F-Col1a1 小鼠，在其他非造血细胞中删除了 FLS 中的 HK2，以及 克霉唑治疗可将 HK2 与线粒体分离，显着降低关节炎的严重程度。 因此，我们将检验线粒体 HK2 是 FLS 表型和 MO 激活的关键调节因子的假设， 这会导致 RA 的关节破坏。 HK2 的鉴定，一种异构体特异性贡献者 RA滑膜组织中细胞葡萄糖代谢升高提供了比整体糖酵解更安全的方法 抑制。 HK2 可以在不损害系统稳态的情况下选择性地靶向 正常细胞中相应的代谢功能作为一种新的组合方法 独立于全身免疫抑制的 RA 关节疾病治疗。