Diamond Nanoparticles Suppress Migration of T98G Glioblastoma Cells by Targeting ECM–Integrin Interactions and Intracellular Signaling, Leading to Extensive Proteome Alterations

Katarzyna Zawadzka, Barbara Wójcik, Malwina Sosnowska-Ławnicka, Marta Kutwin, Sławomir Jaworski, Agnieszka Ostrowska, Michał Pruchniewski, Mateusz Wierzbicki Department of Nanobiotechnology, Institute of Biology, Warsaw University of Life Sciences, Warsaw, 02-786, PolandCorrespondence: Mateusz Wierzbicki, Department of Nanobiotechnology, Institute of Biology, Warsaw University of Life Sciences, Bldg. 23, rm. 133, 8 Ciszewskiego Str., Warsaw, 02-786, Poland, Tel +48 22 593 66 76, Email [email protected]: Glioblastoma (GBM) is a highly heterogeneous and aggressive tumor characterized by rapid growth and therapy resistance. The dynamic interactions of tumor cells with the extracellular matrix (ECM) contribute to treatment inefficacy. While diamond nanoparticles (NDs) are emerging as potential antitumor agents, their mechanisms remain incompletely understood. In this study, we investigated spherical NDs with distinct surface compositions and hydrocolloidal stability and their role in regulating crucial cellular processes in T98G glioblastoma cells.Methods: Two types of detonation diamond nanoparticles (NDs) were characterized using TEM imaging and hydrocolloidal stability assessment in various diluents. Their effects on T98G glioblastoma cells were examined through SEM imaging, cytotoxicity assays, monitoring of spontaneous and collective migration, and early adhesion examination combined with an extensive integrin-blocking panel. Furthermore, characterization via mass spectrometry provided deeper insight into how physicochemical differences between the two NDs types modulate glioblastoma microenvironment and cell responses.Results: NDs were observed to be both intensively internalized by cells and bound to cell membrane, influencing cellular interactions with the extracellular environment. NDs significantly reduced T98G glioblastoma cell migration within 48 hours and impaired early adhesion by effectively blocking α/β integrins. Modified NDs (NDM) demonstrated enhanced hydrocolloidal stability and stronger integrin blocking efficiency. Proteomic analysis revealed that NDs downregulated proteins involved in RNA processing, splicing, and translation while upregulating ECM-related proteins, which profile changed depending on the NDs type.Conclusion: These findings suggest that NDs with distinct surface properties may interact with multiple surface receptors, independently modulate intracellular signaling pathways, and remodel the tumor microenvironment by altering ECM protein composition, positioning them as versatile, multi-targeting agents with antitumor potential.Keywords: glioblastoma, diamond nanoparticles, proteomics, migration, integrins, T98G