Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Heidelberglaan 100, 3508 GA, Utrecht, The Netherlands.
Lowy Cancer Research Centre, Children's Cancer Institute, UNSW Sydney, Sydney, NSW, Australia.
Sci Rep. 2024 Sep 17;14(1):21710. doi: 10.1038/s41598-024-72492-y.
The prognosis of patients with high-risk neuroblastoma remains poor, partly due to inadequate immune recognition of the tumor. Neuroblastomas display extremely low surface MHC-I, preventing recognition by cytotoxic T lymphocytes (CTLs) and contributing to an immunosuppressive tumor microenvironment. Glycogen synthase kinase-3 beta (GSK-3β) is involved in pathways that may affect the MHC-I antigen processing and presentation pathway. We proposed that therapeutic inhibition of GSK-3β might improve the surface display of MHC-I molecules on neuroblastoma cells, and therefore tested if targeting of GSK-3β using the inhibitor 9-ING-41 (Elraglusib) improves MHC-I-mediated CTL recognition. We analyzed mRNA expression data of neuroblastoma tumor datasets and found that non-MYCN-amplified neuroblastomas express higher GSK-3β levels than MYCN-amplified tumors. In non-MYCN-amplified cells SH-SY5Y, SK-N-AS and SK-N-SH 9-ING-41 treatment enhanced MHC-I surface display and the expression levels of a subset of genes involved in MHC-I antigen processing and presentation. Further, 9-ING-41 treatment triggered increased STAT1 pathway activation, upstream of antigen presentation pathways in two of the three non-MYCN-amplified cell lines. Finally, in co-culture experiments with CD8 + T cells, 9-ING-41 improved immune recognition of the neuroblastoma cells, as evidenced by augmented T-cell activation marker levels and T-cell proliferation, which was further enhanced by PD-1 immune checkpoint inhibition. Our preclinical study provides experimental support to further explore the GSK-3β inhibitor 9-ING-41 as an immunomodulatory agent to increase tumor immune recognition in neuroblastoma.
高危神经母细胞瘤患者的预后仍然较差，部分原因是肿瘤的免疫识别不足。神经母细胞瘤表面 MHC-I 极低，阻止了细胞毒性 T 淋巴细胞 (CTL) 的识别，并导致免疫抑制肿瘤微环境。糖原合成酶激酶-3β (GSK-3β) 参与可能影响 MHC-I 抗原加工和呈递途径的途径。我们提出，抑制 GSK-3β 可能会改善神经母细胞瘤细胞表面 MHC-I 分子的表达，因此我们测试了使用抑制剂 9-ING-41（Elraglusib）靶向 GSK-3β 是否会改善 MHC-I 介导的 CTL 识别。我们分析了神经母细胞瘤肿瘤数据集的 mRNA 表达数据，发现非 MYCN 扩增的神经母细胞瘤比 MYCN 扩增的肿瘤表达更高水平的 GSK-3β。在非 MYCN 扩增的细胞 SH-SY5Y、SK-N-AS 和 SK-N-SH 中，9-ING-41 治疗增强了 MHC-I 表面表达和参与 MHC-I 抗原加工和呈递的一组基因的表达水平。此外，在两种非 MYCN 扩增细胞系中的两种中，9-ING-41 治疗触发了抗原呈递途径上游的 STAT1 途径激活。最后，在与 CD8+T 细胞的共培养实验中，9-ING-41 增强了神经母细胞瘤细胞的免疫识别，这表现为 T 细胞激活标志物水平和 T 细胞增殖的增加，而 PD-1 免疫检查点抑制进一步增强了这种增加。我们的临床前研究为进一步探索 GSK-3β 抑制剂 9-ING-41 作为免疫调节剂以增加神经母细胞瘤中的肿瘤免疫识别提供了实验支持。
