Institut Cochin, INSERM U1016, CNRS UMR 8104, LabEx INFLAMEX, Université Paris Descartes, Sorbonne Paris Cité, Paris, France; Pôle de Médecine Interne, Centre de Référence pour les vascularites nécrosantes et la sclérodermie systémique, Hôpital Cochin, Assistance Publique-Hôpitaux de Paris (AP-HP), Paris, France.
Service de Médecine Interne A, CHU Dupuytren, Limoges, France; Laboratoire d'Immunologie, EA3842, Faculté de médecine, Limoges, France.
Autoimmun Rev. 2017 Apr;16(4):398-406. doi: 10.1016/j.autrev.2017.02.006. Epub 2017 Feb 14.
The pathophysiology of giant cell arteritis (GCA) and the mechanisms underlying vascular remodeling, are poorly understood. We aimed to compare vascular smooth muscle cells (VSMCs) from patients with GCA and controls by a proteomic and gene expression profile approach and to identify the signaling pathways involved in proliferation.
VSMCs were cultured from temporal artery biopsies (TABs) from patients with biopsy-proven GCA (TAB-GCA), biopsy-negative GCA (TAB-GCA), and diagnosis other than GCA (GCA-control). VSMCs from normal human aorta (HAoSMC) were used as controls. 2D-differential in-gel electrophoresis and Affymetrix chips were used to compare proteomes and gene expression profiles of VSMCs. Proliferation was assessed by BrdU incorporation assay. TAB-GCA and GCA-control TABs underwent immunohistochemistry staining for endothelin-1 (ET-1) and receptors ETR and ETR.
We identified 16, 30 and 2 protein spots differentially expressed between TAB-GCA and GCA-control VSMCs, TAB-GCA and TAB-GCA VSMCs and TAB-GCA and GCA-control VSMCs, respectively (fold change ≥1.5 and p≤0.05). Among the 153 proteins differentially expressed between TAB-GCA and HAoSMC VSMCs, many were linked with ET-1. Genes differentially expressed between TAB-GCA and GCA-control VSMCs were involved in proliferation. ET-1 was identified as a link between genes of interest. Proliferation was reduced for TAB-GCA VSMCs on treatment with the endothelin antagonist macitentan and its active metabolite. Patients showing transmural expression of ET-1 in temporal artery lesions received a significantly higher glucocorticoid daily dose after 6-month follow-up.
Inhibiting the proliferation with macitentan, combined with glucocorticoids, might be a promising therapeutic approach for patients with GCA.
巨细胞动脉炎（GCA）的病理生理学和血管重塑的机制尚不清楚。我们旨在通过蛋白质组学和基因表达谱分析比较 GCA 患者和对照的血管平滑肌细胞（VSMC），并确定参与增殖的信号通路。
从经活检证实的 GCA（TAB-GCA）、活检阴性的 GCA（TAB-GCA）和非 GCA 诊断的患者的颞动脉活检（TAB）中培养 VSMC，将正常人类主动脉（HAoSMC）中的 VSMC 作为对照。使用 2D 差异凝胶电泳和 Affymetrix 芯片比较 VSMC 的蛋白质组和基因表达谱。通过 BrdU 掺入测定评估增殖。对 TAB-GCA 和 GCA-control TAB 进行内皮素-1（ET-1）和受体 ETR 和 ETR 的免疫组织化学染色。
我们分别在 TAB-GCA 和 GCA-control VSMC、TAB-GCA 和 TAB-GCA VSMC 以及 TAB-GCA 和 GCA-control VSMC 之间鉴定出 16、30 和 2 个差异表达的蛋白质斑点（倍数变化≥1.5 和 p≤0.05）。在 TAB-GCA 和 HAoSMC VSMC 之间差异表达的 153 种蛋白质中，许多与 ET-1 有关。在 TAB-GCA 和 GCA-control VSMC 之间差异表达的基因参与增殖。ET-1 被确定为感兴趣基因之间的联系。用内皮素拮抗剂 macitentan 及其活性代谢物治疗 TAB-GCA VSMC 后，其增殖减少。在 6 个月随访后，接受糖皮质激素治疗的患者，其颞动脉病变中 ET-1 的跨壁表达明显增加。
用 macitentan 抑制增殖，结合糖皮质激素，可能是 GCA 患者有前途的治疗方法。
