Screening anticancer peptides performance in organotypic prostate tumor‐stroma 3D models

Prostate cancer (PCa) poses a significant concern in the realm of cancer, representing a continuous challenge for the scientific community to discover effective therapeutic approaches. Among emerging strategies, anticancer peptides have garnered attention for their potential to disrupt protein-protein interactions. Targeting protein phosphatase 1 (PP1) complexes through PP1-disrupting peptides holds promise for selectively impeding critical pathways in the development and progression of cancer. In this context, CAVPENET peptide was designed to specifically target and disrupt the complex formed between PP1 and caveolin-1, a contributor to the progression of PCa. Previous research has revealed that CAVPENET inhibits the growth of PCa cell 2D monolayers, primarily by modulating PP1 activity. In this study, we developed an increasing physiomimetic human 3D PCa/prostate cancer-associated fibroblast heterotypic spheroid model to evaluate the tumor-suppressive activity of CAVPENET peptide in a more relevant preclinical context. Our findings reveal the formation of morphologically well-defined tumor microtissues that increase their size and cellular density over time, characteristics of in vivo tumors. Upon incubation with CAVPENET, PCa spheroids exhibited decreased growth and viability. In contrast, CAVPENET treatment (20 μM) did not influence CAFs monotypic spheroids growth. In conclusion, our results underscore the relevance of employing 3D PCa-stroma heterotypic models for evaluating anticancer therapeutics and emphasize the therapeutic potential of CAVPENET peptide for PCa.