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Inhibition of CHI3L1 decreases N-cadherin and VCAM-1 levels in glioblastoma
Agnieszka Rusak1, Marlena Gąsior-Głogowska2, Azzurra Sargenti3
1Division of Histology and Embryology, Department of Human Morphology and Embryology, Wroclaw Medical University, T. Chałubińskiego 6a, Wroclaw, 50-368, Poland. agnieszka.rusak@umw.edu.pl.
Background:
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.
Insights
Inhibiting the protein CHI3L1 shows potential in reducing glioblastoma (GBM) invasiveness. This study used a 3D GBM spheroid model to explore CHI3L1 inhibition
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- The protein CHI3L1 promotes cancer progression via angiogenesis, invasion, and immunomodulation.
- High CHI3L1 expression and angiogenesis are characteristic of glioblastoma multiforme (GBM).
- CHI3L1 inhibition presents a potential therapeutic strategy for precision medicine in GBM.
Purpose of the Study:
- To investigate the effect of CHI3L1 inhibition on GBM invasiveness using a 3D spheroid model.
- To analyze the interaction between CHI3L1 and the inhibitor G721-0282.
- To assess changes in key proteins involved in GBM cell adhesion and the tumor microenvironment.
Main Methods:
- In silico molecular docking and dynamics simulations to study CHI3L1-inhibitor interactions.
- Infrared spectroscopy and label-free physical cytometry for inhibitor uptake analysis.
- Immunohistochemistry, Western blot, and ddPCR to evaluate changes in E-cadherin, N-cadherin, VE-cadherin, VCAM-1, and EGFR in U-87 MG cells and GBM spheroids.
Main Results:
- A direct interaction between CHI3L1 and the inhibitor G721-0282 was confirmed.
- G721-0282 treatment reduced N-cadherin and VCAM-1 levels in 3D GBM spheroids.
- Observed changes in protein expression differed between 2D cell cultures and 3D spheroids.
Conclusions:
- CHI3L1 inhibition demonstrates potential in reducing GBM tumor invasiveness.
- The 3D GBM spheroid model is crucial for studying alterations in membrane proteins and the tumor microenvironment.
- Targeting CHI3L1 offers a promising avenue for novel GBM therapies.
