Rusak Agnieszka, Gąsior-Głogowska Marlena, Sargenti Azzurra, Krzyżak Edward, Kotowski Krzysztof, Mrozowska Monika, Górnicki Tomasz, Kujawa Krzysztof, Dzięgiel Piotr
Division of Histology and Embryology, Department of Human Morphology and Embryology, Wroclaw Medical University, T. Chałubińskiego 6a, Wroclaw, 50-368, Poland.
Department of Biomedical Engineering, Faculty of Fundamental Problems of Technology, Wroclaw University of Science and Technology, 27, S. Wyspiańskiego, Wroclaw, 50-370, Poland.
Pharmacol Rep. 2025 Feb;77(1):210-228. doi: 10.1007/s43440-024-00677-3. Epub 2024 Nov 14.
The protein CHI3L1 contributes to cancer development by several mechanisms, including stimulation of angiogenesis and invasion as well as immunomodulatory effects. These properties make it a potential target for the development of targeted therapies in precision medicine. In this context, the particular potential of CHI3L1 inhibition could be considered in glioblastoma multiforme (GBM), whose tumors exhibit high levels of angiogenesis and increased CHI3L1 expression. This study aims to investigate whether inhibition of CHI3L1 in spheroids used as a GBM model affects the mechanisms of invasiveness; METHODS: We analyzed the interactions between CHI3L1 and the inhibitor G721-0282 in molecular docking and molecular dynamics (in silico) and infrared spectroscopy. Uptake of G721-0282 in GBM spheroids was measured using a label-free physical cytometer. Changes in E-, N- and VE-cadherins, VCAM-1, and EGFR were analyzed by immunohistochemical reactions, Western blot, and ddPCR methods in U-87 MG cells and GBM spheroids consisting of U-87 MG glioblastoma cells, HMEC-1 endothelial cells and macrophages; RESULTS: A direct interaction between CHI3L1 and G721-0282 was confirmed. G721-0282 decreased N-cadherins and VCAM-1 in GBM spheroids, but the changes in the 2D model of U-87 MG glioblastoma cells were different; CONCLUSION: Inhibition of CHI3L1 has the potential to reduce the invasiveness of GBM tumors. The 3D model of GBM spheroids is of great significance for investigating changes in membrane proteins and the tumor microenvironment.
蛋白质CHI3L1通过多种机制促进癌症发展，包括刺激血管生成和侵袭以及免疫调节作用。这些特性使其成为精准医学中靶向治疗开发的潜在靶点。在这种背景下，CHI3L1抑制的特殊潜力可在多形性胶质母细胞瘤（GBM）中加以考虑，该肿瘤表现出高水平的血管生成和CHI3L1表达增加。本研究旨在调查在用作GBM模型的球体中抑制CHI3L1是否会影响侵袭机制；方法：我们在分子对接和分子动力学（计算机模拟）以及红外光谱中分析了CHI3L1与抑制剂G721-0282之间的相互作用。使用无标记物理细胞仪测量GBM球体对G721-0282的摄取。通过免疫组化反应、蛋白质免疫印迹和ddPCR方法分析了U-87 MG细胞以及由U-87 MG胶质母细胞瘤细胞、HMEC-1内皮细胞和巨噬细胞组成的GBM球体中E-钙黏蛋白、N-钙黏蛋白和血管内皮钙黏蛋白、血管细胞黏附分子-1（VCAM-1）和表皮生长因子受体（EGFR）的变化；结果：证实了CHI3L1与G721-0282之间存在直接相互作用。G721-0282降低了GBM球体中的N-钙黏蛋白和VCAM-1，但U-87 MG胶质母细胞瘤细胞二维模型中的变化有所不同；结论：抑制CHI3L1有可能降低GBM肿瘤的侵袭性。GBM球体的三维模型对于研究膜蛋白和肿瘤微环境的变化具有重要意义。
