Lee Kong Chian School of Medicine, Nanyang Technological University Singapore, Singapore, Singapore.
Interdisciplinary Graduate Programme, NTU Institute for Health Technologies (HealthTech NTU), Nanyang Technological University Singapore, Singapore, Singapore.
Front Immunol. 2021 Dec 17;12:680071. doi: 10.3389/fimmu.2021.680071. eCollection 2021.
The trafficking of T-cells through peripheral tissues and into afferent lymphatic vessels is essential for immune surveillance and an adaptive immune response. Glycogen synthase kinase 3β (GSK3β) is a serine/threonine kinase and regulates numerous cell/tissue-specific functions, including cell survival, metabolism, and differentiation. Here, we report a crucial involvement of GSK3β in T-cell motility. Inhibition of GSK3β by CHIR-99021 or siRNA-mediated knockdown augmented the migratory behavior of human T-lymphocytes stimulated an engagement of the T-cell integrin LFA-1 with its ligand ICAM-1. Proteomics and protein network analysis revealed ongoing interactions among GSK3β, the surface receptor Notch1 and the cytoskeletal regulator CRMP2. LFA-1 stimulation in T-cells reduced Notch1-dependent GSK3β activity by inducing phosphorylation at Ser9 and its nuclear translocation accompanied by the cleaved Notch1 intracellular domain and decreased GSK3β-CRMP2 association. LFA-1-induced or pharmacologic inhibition of GSK3β in T-cells diminished CRMP2 phosphorylation at Thr514. Although substantial amounts of CRMP2 were localized to the microtubule-organizing center in resting T-cells, this colocalization of CRMP2 was lost following LFA-1 stimulation. Moreover, the migratory advantage conferred by GSK3β inhibition in T-cells by CHIR-99021 was lost when CRMP2 expression was knocked-down by siRNA-induced gene silencing. We therefore conclude that GSK3β controls T-cell motility through interactions with CRMP2 and Notch1, which has important implications in adaptive immunity, T-cell mediated diseases and LFA-1-targeted therapies.
T 细胞通过外周组织转移到输入淋巴管对于免疫监视和适应性免疫反应至关重要。糖原合酶激酶 3β（GSK3β）是一种丝氨酸/苏氨酸激酶，调节许多细胞/组织特异性功能，包括细胞存活、代谢和分化。在这里，我们报告了 GSK3β 参与 T 细胞迁移的关键作用。用 CHIR-99021 或 siRNA 介导的敲低抑制 GSK3β 增强了人类 T 淋巴细胞的迁移行为，刺激了 T 细胞整合素 LFA-1 与其配体 ICAM-1 的结合。蛋白质组学和蛋白质网络分析揭示了 GSK3β、表面受体 Notch1 和细胞骨架调节剂 CRMP2 之间持续的相互作用。T 细胞中 LFA-1 的刺激通过诱导 Ser9 磷酸化及其核易位，并伴有裂解的 Notch1 细胞内结构域和减少 GSK3β-CRMP2 相互作用，降低了 Notch1 依赖性 GSK3β 活性。T 细胞中 LFA-1 诱导或药物抑制 GSK3β 可减少 CRMP2 在 Thr514 处的磷酸化。尽管在静止的 T 细胞中，大量的 CRMP2 定位于微管组织中心，但在 LFA-1 刺激后，这种 CRMP2 的共定位消失了。此外，当通过 siRNA 诱导的基因沉默敲低 CRMP2 表达时，CHIR-99021 抑制 GSK3β 在 T 细胞中赋予的迁移优势丧失。因此，我们得出结论，GSK3β 通过与 CRMP2 和 Notch1 的相互作用来控制 T 细胞的迁移，这对适应性免疫、T 细胞介导的疾病和 LFA-1 靶向治疗具有重要意义。
