Nanobody-based immunotoxins: A precision tool in the treatment of solid tumors

Solid tumors, the main cause of cancer-related death, represent a significant therapeutic challenge due to the high-density microenvironment and intolerance to conventional treatments. Nanobody-based immunotoxins (NbITs) are an exciting candidate, combining the ultimate specificity of nanobodies (single-domain antibody fragments of camelid antibodies) and detrimental effects of the toxin. These nanobodies are small (one-tenth of conventional antibody size), thermostable with high specificity, high antigen binding affinity which give it the ability to penetrate into solid tumors. Specific delivery to tumor cells is achieved through conjugating nanobodies with cytotoxic agents of bacterial origin or synthetic drugs. This phenomenon is initially attracted to the cells by the antigen-antibody interaction that is further enhanced by receptor-mediated internalization and cytotoxic payload release that subdues essential cellular processes and, as a consequence, damages the cells. This review discusses the mechanisms that underlie the effectiveness of NbITs, such as tumor antigen recognition, toxin release, and cellular signaling pathways elicited by the internalized toxins. We also discuss the application of NbITs in treating cancers such as HER2-positive breast cancer and EGFR-overexpressing lung cancer, and other cancers, highlighting their ability to address limitations of conventional therapies. Key challenges in NbIT development, including stability, immunogenicity, and efficient delivery, are critically evaluated. Current advances such as the creation of bispecific nanobody constructs, optimization of linker strategies, as well as the incorporation of nanoparticle-based delivery systems are maximizing the therapeutic potential of these molecules. This review synthesizes recent progress and addresses current obstacles in NbIT development, showcasing their transformative potential as a targeted therapeutic approach for solid tumors. It also covers future opportunities to develop and advance this emerging treatment strategy.