Design of TNFR Peptide Agonists for Inducing Receptors Oligomerization and Cell Apoptosis

Abstract

TNFRs within the tumor necrosis factor receptor superfamily represent promising therapeutic targets for cancer treatment, particularly for chemotherapy-resistant cells capable of p53-independent apoptosis induction. We developed peptide agonists utilizing β-sheet structures from FasL and TRAIL as scaffolds to induce 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, validating 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.