An Antibody Targeting Fibroblast Activation Protein Simultaneously Fused to Interleukin-2 and Tumor Necrosis Factor Selectively Localizes to Neoplastic Lesions

• Published in: Antibodies (Basel) Vol. 12; no. 2; p. 29
• Main Authors: Prodi, Eleonora, Comacchio, Claudia, Gilardoni, Ettore, Di Nitto, Cesare, Puca, Emanuele, Neri, Dario, De Luca, Roberto
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 14.04.2023; MDPI
• Subjects: Antibodies; antibody engineering; antibody–cytokine fusion proteins; Anticancer properties; Antigens; Antimitotic agents; Antineoplastic agents; Antitumor activity; Biological activity; Cancer; Cells; Chromatography; Cytokines; Fibroblast activation protein; Fibroblasts; Fusion protein; Health aspects; Immune response; Immune system; In vivo methods and tests; Interleukin 2; Localization; Payloads; Physiological aspects; Proteins; Testing; Trimers; Tumor necrosis factor; Tumor necrosis factor-TNF; Tumors; Viral antibodies; Italy; United States > US; antibody engineering; interleukin-2; antibody–cytokine fusion proteins; tumor necrosis factor; fibroblast activation protein
• ISSN: 2073-4468; 2073-4468
• DOI: 10.3390/antib12020029

The delivery of specific cytokine payloads to a neoplastic environment employing antibodies able to selectively accumulate at the tumor site represents an attractive strategy to stimulate an immune response to cancer. Whilst conventional antibody-cytokine fusions based on a single payload have shown potent anticancer activity, the concomitant delivery of two cytokine payloads may further improve the therapeutic outcome as the immune system typically adopts multiple signals to reinforce an antitumor strategy. We here describe a potency-matched dual-cytokine antibody fusion protein containing a tumor-targeting antibody fragment specific to human fibroblast activation protein (FAP), simultaneously linked to both interleukin-2 (IL2) and a tumor necrosis factor (TNF) mutant. The resulting fusion protein, termed IL2-7NP2-TNF , formed stable non-covalent trimers driven by the interaction of the tumor necrosis factor subunits. Both cytokine payloads retained their biological activity within the fusion protein, as shown by in vitro cellular assays. The tumor-targeting properties and the anticancer activity of IL2-7NP2-TNF were investigated in vivo in immunocompromised mice bearing SKRC52 cells transduced with human FAP. The fusion protein preferentially localized to the cancer site and induced partial tumor retardation.