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: 10410487
• 负责人: Monica Guma
• 金额: $ 34.41万
• 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 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10410487
• 关键词: Address; Adult; Affect; Apoptosis; Arthritis; Binding; Cartilage; Cells; Cellular Metabolic Process; Color; Combined Modality Therapy; Confocal Microscopy; Cytosol; Data; Development; Disease; Endoplasmic Reticulum; Enzymes; Fibroblasts; Glucose Transporter; Glycolysis; Glycolysis Inhibition; Growth; Hexokinase 2; Homeostasis; Hypoxia; Immunosuppression; Impairment; In Vitro; Inflammation; Inflammatory; Inflammatory Arthritis; Injections; Interleukin-6; Intra-Articular Injections; Joints; Knee; Lesion; Macrophage Activation; Membrane; Membrane Proteins; Metabolic; Metabolism; Miconazole; Mitochondria; Molecular; Mus; Normal Cell; Pathogenesis; Pathway interactions; Patients; Peptides; Pharmaceutical Preparations; Phenotype; Phosphorylation; Phosphotransferases; Play; Population; Process; Protein Isoforms; Rheumatism; Rheumatoid Arthritis; Risk; Role; Sampling; Severities; Signal Pathway; Stromal Cells; Synovial Cell; Synovial Fluid; Synovial Membrane; Synovitis; Testing; Therapeutic; Thick; Tissues; Up-Regulation; Voltage-Dependent Anion Channel; Work; adenoviral-mediated; arthropathies; bone; bone cell; cell motility; cell type; extracellular; glucose metabolism; hexokinase; improved; in vivo; insight; joint destruction; joint inflammation; knock-down; macrophage; migration; mutant; novel; novel therapeutics; overexpression; peptide drug; public health relevance; voltage-dependent anion channel 2

ABSTRACT Targeting Hexokinase 2 in Rheumatoid Arthritis (changes are underlined) Hexokinases (HKs) catalyze the first committed step in glucose metabolism. HK2 constitutes the principal inducible isoform and has a restricted distribution of expression in normal adult tissues. Cell populations with increased glycolysis and HK2 expression have a powerful growth advantage. HK2 localizes also at mitochondria, and its interaction increases glucose metabolism and protects mitochondria against apoptosis. Thus, mitochondrial HK2 translocation promotes an activated phenotype in several cell types. Fibroblast like synoviocytes (FLS) and macrophages (MO) are a key component of rheumatoid arthritis (RA) inflamed synovium and contribute to the initiation and perpetuation of destructive joint inflammation. FLS from patients with RA display unique aggressive features, which are autonomous and vertically transmitted. MO are also critical in the pathogenesis of RA. The increase in the number of sublining MO in the synovium is an early hallmark of active rheumatic disease, and high numbers of MO are a prominent feature of inflammatory lesions. Of note, a critical role of glucose metabolism in both activated FLS and MO, have been highlighted by our recent work among others. 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.

摘要 靶向Hexokinase2治疗类风湿关节炎(带下划线的变化) 己糖激酶(HKS)催化葡萄糖代谢的第一步。港币2为主要 可诱导的异构体，在正常成人组织中的表达分布有限。细胞群体与 糖酵解增加和HK2表达具有强大的生长优势。HK2的本地化也在 线粒体及其相互作用可促进葡萄糖代谢并保护线粒体免受细胞凋亡的影响。 因此，线粒体HK2易位促进了几种细胞类型的激活表型。 成纤维细胞样滑膜细胞(FLS)和巨噬细胞(MO)是类风湿关节炎(RA)的重要组成部分 滑膜发炎，并导致破坏性关节炎症的开始和持续。FLS来自 RA患者表现出独特的攻击性特征，这些特征是自主的和垂直传播的。莫都是 在类风湿关节炎的发病机制中也起重要作用。滑膜中次衬里MO数量的增加是早期的 活动性风湿病的标志和大量的MO是炎症性疾病的显著特征 损伤。值得注意的是，糖代谢在激活的FLS和MO中的关键作用已经被强调 在其他方面，我们最近的工作。 我们的初步数据表明，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滑膜中观察到，而在OA滑膜中不表达 样本。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，HK2过表达可增加FLS的侵袭率和移行率。值得注意的是乳酸和PLOD2， 参与细胞迁移和侵袭的基因在HK2表达后上调。上调对 胞外乳酸也提示代谢转向加速糖酵解代谢。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，一种能够 使HK2与线粒体分离，损害IL-6的分泌。重要的是，腺病毒介导的表达 关节内注射HK2引起的膝关节滑膜增厚，当 关节腔内注射HK2ΔN。最后，HK2F/F-Col1a1小鼠，它删除了FLS中的HK2和其他非 造血细胞，以及克霉唑的治疗，它将HK2从线粒体中分离出来，显著地 降低了关节炎的严重程度。因此，我们将检验线粒体HK2是FLS关键调节因子的假设 表型和MO激活，这有助于RA的联合破坏。香港2号、香港2号和香港2号 类风湿关节炎滑膜组织中细胞葡萄糖代谢升高的异构体特异性贡献者提供了一个更安全的 比整体糖酵解抑制方法更有效。香港二号可以在不妥协的情况下选择性地成为目标 作为一种新的补充因素，正常细胞的系统内稳态或相应的代谢功能 不依赖全身免疫抑制的RA关节疾病综合治疗方法。