Phosphorylated Platelet-Derived Growth Factor Receptor-Positive Cells With Anti-apoptotic Properties Accumulate in the Synovium of Patients With Rheumatoid Arthritis
Phosphorylated Platelet-Derived Growth Factor Receptor-Positive Cells With Anti-apoptotic Properties Accumulate in the Synovium of Patients With Rheumatoid Arthritis
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DOI：
10.3389/fimmu.2019.00241
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发表时间：
2019-02-15
影响因子：
7.3
通讯作者：
Saito-Chikenji, Takako
中科院分区：
文献类型：
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作者：
Matsumura, Takashi;Saito, Yuki;Saito-Chikenji, Takako
风湿性关节炎（RA）是由滑膜炎症引起的自身免疫性疾病，其特征在于破坏骨和软骨的慢性多关节炎。类风湿关节炎患者滑膜中的成纤维细胞样滑膜细胞（FLS）可通过产生基质金属蛋白酶和NF-κ B配体的受体激活剂等蛋白质促进软骨和骨的破坏，因此代表了类风湿关节炎的重要治疗靶点。FLS有几种表型，这取决于细胞表面蛋白和粘附因子的表达。识别与不同表型相关的细胞功能和控制它们的方法被认为是开发RA治疗策略的关键。在这项研究中，滑膜组织收集自RA患者和由于韧带损伤或骨折而需要手术的对照受试者。免疫组织学分析用于研究磷酸化血小板衍生生长因子受体-α β（pPDGFR α β）和钙粘蛋白-11（CDH 11）表达的阳性率，并评估每种细胞表型的血小板减少相关标志物。接下来，在体外分离FLS，并且除了PDGF和转化生长因子（2GF）的组合之外，还用肿瘤坏死因子-α（TNF-α）刺激，以研究pPDGFR α β和CDH 11表达以及TNF和细胞周期蛋白依赖性激酶（CDK）4/6的抑制对FLS的影响。免疫组织学分析显示，RA患者的内衬下层（SL）中有很大比例的pPDGFR α β + CDH 11-细胞。这些细胞表现出B细胞淋巴瘤-2表达增加，TNF受体-1表达减少，对细胞死亡的抗性和异常增殖，表明有在滑膜中积聚的趋势。此外，在体外，FLS的2GF刺激降低，而2GF + TNF刺激增加pPDGFR α β/CDH 11比率。假设用2GF + TNF刺激的FLS会在RA体内蓄积，我们确定了TNF和CDK 4/6抑制剂的治疗效果。TNF抑制剂降低pPDGFR α β/CDH 11比率，而CDK 4/6抑制剂抑制细胞增殖。然而，通过组合两种药物未观察到协同效应。我们观察到RA滑膜SL中pPDGFR α β + CDH 11-细胞的增加以及这些细胞在滑膜中的积累。我们发现TNF抑制剂抑制FLS活性，CDK 4/6抑制剂降低细胞增殖。
Rheumatoid arthritis (RA) is an autoimmune disease caused by inflammation of the synovium and characterized by chronic polyarthritis that destroys bone and cartilage. Fibroblast-like synoviocytes (FLSs) in the synovium of patients with RA can promote cartilage and bone destruction by producing proteins such as matrix metalloproteinases and receptor activator of NF-kappa B ligand, thereby representing an important therapeutic target for RA. FLSs have several phenotypes depending on which cell surface proteins and adhesion factors are expressed. Identifying the cellular functions associated with different phenotypes and methods of controlling them are considered essential for developing therapeutic strategies for RA. In this study, synovial tissue was collected from patients with RA and control subjects who required surgery due to ligament injury or fracture. Immunohistological analysis was used to investigate the rates of positivity for phosphorylated platelet-derived growth factor receptor-alpha beta(pPDGFR alpha beta) and cadherin-11 (CDH11) expression, and apoptosis-related markers were assessed for each cell phenotype. Next, FLSs were isolated in vitro and stimulated with tumor necrosis factor-alpha (TNF-alpha) in addition to a combination of PDGF and transforming growth factor (2GF) to investigate pPDGFR alpha beta and CDH11 expression and the effects of the inhibition of TNF and cyclin-dependent kinase (CDK) 4/6 on FLSs. Immunohistological analysis showed a large percentage of pPDGFR alpha beta + CDH11-cells in the sub-lining layer (SL) of patients with RA. These cells exhibited increased B-cell lymphoma-2 expression, reduced TNF receptor-1 expression, resistance to cell death, and abnormal proliferation, suggesting a tendency to accumulate in the synovium. Further, in vitro 2GF stimulation of FLSs lowered, whereas 2GF + TNF stimulation increased the pPDGFR alpha beta/CDH11 ratio. Hypothesizing that FLSs stimulated with 2GF + TNF would accumulate in vivo in RA, we determined the therapeutic effects of TNF and CDK4/6 inhibitors. The TNF inhibitor lowered the pPDGFR alpha beta/CDH11 ratio, whereas the CDK4/6 inhibitor suppressed cell proliferation. However, a synergistic effect was not observed by combining both the drugs. We observed an increase in pPDGFR alpha beta+CDH11-cells in the SL of the RA synovium and accumulation of these cells in the synovium. We found that the TNF inhibitor suppressed FLS activity and the CDK4/6 inhibitor reduced cell proliferation.