Engineering nano-clustered multivalent agonists to cross-link TNF receptors for cancer therapy

Tumor necrosis factor receptors (TNFRs) are promising targets for cancer therapy. However, activating their downstream signaling requires cross-linking of TNFRs. Herein, to devise strong agonists of TNFRs, ligands targeting TNFRs, such as OX40L and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), were fused with a multivalent protein scaffold (MV) to prepare multivalent agonists for cross-linking TNFRs. The nano-clustered multivalent-OX40L (MV-OX40L) and MV-TRAIL could promote T cell activation and directly induce tumor cell apoptosis. Moreover, to develop a universal nano-adaptor for the rapid preparation of multivalent agonists of different TNFRs, the Fc receptor that could immobilize antibodies was fused with MV to prepare MV-FcR, which could multimerize commercial agonist antibodies targeting TNFRs, such as anti-OX40 antibody (& alpha;OX40). Simply incubating & alpha;OX40 with MV-FcR could prepare MV-& alpha;OX40 to enhance its antitumor efficacy. In addition, MV-FcR could multimerize with other therapeutic antibodies, such as anti-PD-L1 antibody, to enhance their valency. This study provides a promising strategy for engineering multivalent antitumor protein drugs.