Growing evidence has unveiled the pathological significance of Tau in many cancers, including the most aggressive and lethal brain tumor glioblastoma multiform (GBM). In this regard, we have recently examined the structure–activity relationship of a new series of seventeen 2-aminothiazole-fused to flavonoid hybrid compounds (TZF) on Tau-overexpressing GBM cells. Here, we evaluated the anticancer activities of the two lead compounds 2 and 9 using multi-cellular spheroids (MCSs) which represent an easy 3D human cell model to mimic GBM organization, physical constraints and drug penetration. The two compounds reduced cell evasion from spheroids up to three times, especially for Tau-expressing cells. As a first step towards a therapeutic approach, we quantified the effects of these compounds on MCS growth using two complementary protocols: single and repeated treatments. A single injection with compound 9 slowed down the growth of MCSs formed with U87 shCTRL cells by 40% at 10 µM. More interestingly, multiple treatment with compound 9 slowed the growth of U87 shCTRL spheroids by 40% at a concentration of 5 µM, supporting the increased bioavailability of compound 9 within MCSs. In conclusion, compound 9 deserves particular attention as promising candidate for specifically targeting Tau-expressing cancers such as GBM.

Here, we present a protocol for achieving high-density peptide and protein presentation on biomaterial surfaces using self-assembling virus-like particles. We describe steps for the design, recombinant production, purification, and quality control of peptide- or protein-displaying particles. We also detail procedures for their use in functionalizing polydimethylsiloxane (PDMS) and other solid substrates. The approach enables multivalent and spatially controlled ligand presentation and is applicable to reproducible cell-material interaction studies and the biofunctionalization of implant-relevant surfaces. For complete details on the use and execution of this protocol, please refer to Maayouf et al.¹.