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.

项目摘要 经过几年的生物疗法和小分子靶向炎症的研究，我们需要一个 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反应者。在其他领域，如肿瘤学 通过代谢重编程来改善免疫治疗的概念是我们真正认为应该 转化为自身免疫性疾病，以补充目前的治疗方法。然而，关于 针对RA的代谢变化。我们希望通过这笔赠款更好地了解这些生物学 代谢途径，以更好地表征其治疗设备的新方法。我们的发现 己糖激酶2（HK 2）活性仅在RA滑膜和不同RA滑膜细胞中增强，提示 在关节中协同代谢重编程，有助于RA的发展和进展。我们 初步数据表明，虽然HK 1在OA和RA滑膜中表达，但HK 2 表达与MO和FLS标志物共定位，并且仅在RA中观察到，而在OA滑膜样品中未观察到。 我们还表明，HK 2调节关键FLS功能，因为HK 2敲低损害FLS侵袭。相反，HK 2 过表达增加FLS侵袭和迁移率。值得注意的是，乳酸盐和PLOD 2参与了 细胞迁移和侵袭在HK 2表达后上调。细胞外乳酸的上调也 表明代谢向加速糖酵解代谢转变。一个缺乏线粒体的HK 2突变体- 结合基序（HK 2 ΔN）逆转了侵袭表型。在MO中，将HK 2从 线粒体，IL-6分泌受损。重要的是，腺病毒介导的HK 2在膝关节内的表达， 关节注射诱导滑膜增厚，当HK 2 ΔN在关节内时，这一点不太明显 注射。最后，在FLS和其他非造血细胞中缺失HK 2的HK 2F/F-Col 1a 1小鼠， 用克霉唑（其使HK 2与线粒体解离）治疗显著降低了关节炎的严重程度。 因此，我们将检验线粒体HK 2是FLS表型和MO激活的关键调节因子的假设， 这有助于RA中的联合破坏。HK 2的鉴定，一种亚型特异性贡献者， RA滑膜组织中细胞葡萄糖代谢的升高提供了比整体糖酵解更安全的方法 抑制作用HK 2可以选择性靶向而不损害全身稳态， 在正常细胞中的相应代谢功能作为组合的新的附加方法 RA关节疾病的治疗不依赖于全身免疫抑制。