A novel nanobody-based immunocytokine of a mutant interleukin-2 as a potential cancer therapeutic

• Published in: AMB Express Vol. 14; no. 1; p. 19
• Main Authors: Beig Parikhani, Arezoo, Dehghan, Rada, Talebkhan, Yeganeh, Bayat, Elham, Biglari, Alireza, Shokrgozar, Mohammad Ali, Ahangari Cohan, Reza, Mirabzadeh, Esmat, Ajdary, Soheila, Behdani, Mahdi
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 09.02.2024; Springer Nature B.V; SpringerOpen
• Subjects: Antibodies; Biocompatibility; Biological activity; Biomedical and Life Sciences; Biotechnology; Cancer; Cancer therapies; CD25 antigen; Cell activation; Cell proliferation; Cytokines; Cytotoxicity; Effector cells; Flow cytometry; Growth factors; Immunocytokine; Immunotherapy; In vivo methods and tests; Interleukin 2; Interleukins; Leukocytes (mononuclear); Life Sciences; Microbial Genetics and Genomics; Microbiology; Mutant IL-2; Mutants; Nanobodies; Nanobody; Original Article; Peripheral blood mononuclear cells; Pharmacokinetics; Proteins; Toxicity; Tumor microenvironment; Tumors; Vascular endothelial growth factor; Vascular endothelial growth factor receptors; VEGFR2; γ-Interferon; Immunocytokine; VEGFR2; Nanobody; Immunotherapy; Mutant IL-2
• ISSN: 2191-0855; 2191-0855
• DOI: 10.1186/s13568-023-01648-2

The immunotherapeutic application of interleukin-2 (IL-2) in cancer treatment is limited by its off-target effects on different cell populations and insufficient activation of anti-tumor effector cells at the site of the tumor upon tolerated doses. Targeting IL-2 to the tumor microenvironment by generating antibody-cytokine fusion proteins (immunocytokine) would be a promising approach to increase efficacy without associated toxicity. In this study, a novel nanobody-based immunocytokine is developed by the fusion of a mutant (m) IL-2 with a decreased affinity toward CD25 to an anti-vascular endothelial growth factor receptor-2 (VEGFR2) specific nanobody, denoted as VGRmIL2-IC. The antigen binding, cell proliferation, IFN-γ-secretion, and cytotoxicity of this new immunocytokine are evaluated and compared to mIL-2 alone. Furthermore, the pharmacokinetic properties are analyzed. Flow cytometry analysis shows that the VGRmIL2-IC molecule can selectively target VEGFR2-positive cells. The results reveal that the immunocytokine is comparable to mIL-2 alone in the stimulation of Primary Peripheral Blood Mononuclear Cells (PBMCs) and cytotoxicity in in vitro conditions. In vivo studies demonstrate improved pharmacokinetic properties of VGRmIL2-IC in comparison to the wild or mutant IL-2 proteins. The results presented here suggest VGRmIL2-IC could be considered a candidate for the treatment of VEGFR2-positive tumors. Key Points The newly developed IL-2- based immunocytokine may target VEGFR2- positive tumors. The fusion of nanobody to IL-2 improved its pharmacokinetics. The biological activity of IL-2 was not affected by its genetically fusion to nanobody.