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: 10161179
• 负责人: Monica Guma
• 金额: $ 6.58万
• 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/10161179
• 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.

项目总结 经过几年对生物疗法和针对炎症的小分子的研究，我们需要一种 不同的策略，以进一步深入了解RA的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应答者。在其他领域，如肿瘤学 代谢再编程以改进免疫治疗的概念是我们真正认为应该是 转化为自身免疫性疾病，以补充目前的治疗方法。然而，关于这方面的数据很少。 针对类风湿关节炎的代谢变化。我们希望通过这笔赠款更好地了解这些生物的生物学 RA的代谢途径，以更好地描述其治疗手段中的一种新方法。我们的发现 己糖激酶2(HK2)的活性只在RA滑膜和不同的RA滑膜细胞中增强，提示 关节内的协作性代谢再编程有助于类风湿关节炎的发展和进展。我们的 初步数据显示，虽然HK1在OA和RA滑膜中都有表达，但HK2在OA和RA滑膜中都有表达 表达与MO和FLS标记物共定位，仅在RA滑膜标本中观察到，而在OA滑膜标本中未观察到。 我们还发现，当HK2基因被敲除后，FLS的侵袭受到损害，HK2对关键的FLS功能起着调节作用。反之，香港2 过度表达会增加FLS的侵袭率和移行率。值得一提的是，乳酸和PLOD2，它们参与了 细胞迁移和侵袭，在HK2表达后上调。细胞外乳酸的上调也 提示代谢向糖酵解代谢加速转变。一个HK2突变体缺乏线粒体- 结合基序(HK2、Δ、N)逆转了侵袭表型。在MO中，使HK2和HK2解离的一种肽 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 关节内注射可引起滑膜增厚，当HK2ΔN为关节内注射时，滑膜增厚不明显 注射过的。最后，HK2F/F-Col1a1小鼠，在其他非造血细胞中删除FLS中的HK2，以及 克霉唑使HK2从线粒体中分离出来，显著降低了关节炎的严重程度。 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， 这导致了RA的联合破坏。同种异构体特异贡献者HK2的鉴定 RA滑膜组织细胞葡萄糖代谢升高提供了一种比整体糖酵解更安全的方法 抑制力。HK2可以在不损害全身动态平衡的情况下选择性地成为靶点，或者 正常细胞的相应代谢功能作为一种新的结合途径 不依赖全身免疫抑制的RA关节疾病的治疗。