Author(s): Romano, Sofia ; Moura, Vera ; Simões, Sérgio ; Moreira, João Nuno ; Gonçalves, João
Date: 2018
Persistent ID: https://hdl.handle.net/10316/108046
Origin: Estudo Geral - Universidade de Coimbra
Subject(s): Antibody-Dependent Cell Cytotoxicity; Antineoplastic Agents, Immunological; Cell Death; Cell Line, Tumor; Humans; Immunoglobulin Fc Fragments; Immunoglobulin G; Immunoglobulin Heavy Chains; Neoplasms; Phosphoproteins; RNA-Binding Proteins; Single-Domain Antibodies
Nucleolin arises as a relevant target for cancer therapy, as it is overexpressed at the surface of cancer and angiogenic endothelial cells thus enabling a dual cellular targeting strategy. Immunotherapeutic strategies, albeit of proven therapeutic relevance, have been scarcely explored against this target. Therefore, this work aimed at engineering an anti-nucleolin VHH-based antibody capable of triggering anticancer immune responses. Herein, anti-nucleolin VHHs have been generated upon grafting F3 peptide-derived nucleolin-binding sequences onto a VHH CDR1 or CDR3. One of these nucleolin-binding CDR3-grafted VHH was subsequently fused to a human IgG1 Fc region, enabling a significant antibody-dependent cell-mediated cytotoxicity (ADCC). The generated anti-nucleolin VHH revealed increased binding and antiproliferative effects against cancer cells, relative to the parental VHH, while the VHH-Fc counterpart presented increased cytotoxicity relative to the corresponding VHH. This VHH-Fc also triggered an ADCC effect, in the nanomolar range, against a nucleolin-overexpressing cancer cell line. This effect was evidenced by a 2 or 1.7-fold increase of cell death, in the presence of PBMCs, relative to the parental VHH-Fc or the VHH counterpart, respectively. Overall, these formats represent the first anti-nucleolin VHHs and the first anti-nucleolin antibody with ADCC activity that have been successfully developed.
FCT- SFRH/BD/51680/2011
