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.

抽象的 类风湿关节炎中的靶向己糖激酶 2（下划线部分为变化） 己糖激酶 (HK) 催化葡萄糖代谢的第一个关键步骤。 HK2 构成主体 诱导型同种型，在正常成人组织中表达分布有限。细胞群 糖酵解和 HK2 表达增加具有强大的生长优势。 HK2 也本地化于 线粒体及其相互作用增加葡萄糖代谢并保护线粒体免于凋亡。 因此，线粒体 HK2 易位促进了多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) 是类风湿性关节炎 (RA) 的关键组成部分 滑膜发炎，并导致破坏性关节炎症的发生和持续。 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。莫是 在 RA 的发病机制中也至关重要。滑膜中下衬MO数量的增加是早期的表现 活动性风湿病的标志，大量 MO 是炎症的一个显着特征 病变。值得注意的是，葡萄糖代谢在激活的 FLS 和 MO 中的关键作用已被强调 我们最近的工作等等。 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 滑膜中未观察到 样品。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 过表达会增加 FLS 侵袭和迁移率。值得注意的是，乳酸和 PLOD2， HK2 表达后上调，参与细胞迁移和侵袭。上调 细胞外乳酸还表明代谢转向加速糖酵解代谢。 HK2突变体 缺乏线粒体结合基序（HK2ΔN）逆转了侵袭表型。在 MO 中，一种肽 将 HK2 与线粒体解离，损害 IL-6 分泌。重要的是，腺病毒介导的表达 HK2通过关节内注射在膝关节中诱导滑膜厚度，当 HK2ΔN关节内注射。最后，HK2F/F-Col1a1 小鼠，它删除了 FLS 中的 HK2 以及其他非 造血细胞和克霉唑治疗，使 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 关键调节因子的假设 表型和 MO 激活，导致 RA 关节破坏。 HK2的识别 RA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更安全的方法 方法比全局糖酵解抑制更有效。 HK2 可以在不受影响的情况下选择性地成为目标 正常细胞中的系统稳态或相应的代谢功能作为一种新的附加功能 独立于全身免疫抑制的 RA 关节疾病联合治疗方法。