Design of TNFR peptide agonists for inducing receptor oligomerization and cell apoptosis

Tumor Necrosis Factor Receptors (TNFRs), members of the tumor necrosis factor receptor superfamily, are promising therapeutic targets for cancer treatment, particularly for chemotherapy-resistant cells capable of inducing p53-independent apoptosis. We developed peptide agonists utilizing β-sheet structures from FasL and TRAIL as scaffolds to induce the oligomerization of CD95 and death receptor 5 (DR5) receptors. Computational design coupled with AlphaFold3 analysis identified four lead peptides: W10 and AP (targeting CD95) and EIA and EPR (targeting DR5). Bio-layer interferometry demonstrated nanomolar to micromolar binding affinities, while circular dichroism revealed conformational transitions upon target engagement. Target specificity was confirmed through colocalization and oligomerization studies, confirming nanocluster formation. These peptides exhibited micromolar IC50 values against colorectal and breast cancer cell lines. Mechanistic investigations revealed enhanced nuclear translocation of NF-κB associated with apoptotic pathways. This ligand-derived scaffold approach introduces a novel strategy for developing TNFR peptide agonists with significant therapeutic potential against treatment-resistant malignancies.