Near-infrared photoimmunotherapy targeting EGFRShedding new light on glioblastoma treatment

Glioblastomas (GBMs) are high-grade brain tumors, differentially driven by alterations (amplification, deletion or missense mutations) in the epidermal growth factor receptor (EGFR), that carry a poor prognosis of just 12-15 months following standard therapy. A combination of interventions targeting tumor-specific cell surface regulators along with convergent downstream signaling pathways may enhance treatment efficacy. Against this background, we investigated a novel photoimmunotherapy approach combining the cytotoxicity of photodynamic therapy with the specificity of immunotherapy. An EGFR-specific affibody (Z(EGFR:03115)) was conjugated to the phthalocyanine dye, IR700DX, which when excited with near-infrared light produces a cytotoxic response. Z(EGFR:03115)-IR700DX EGFR-specific binding was confirmed by flow cytometry and confocal microscopy. The conjugate showed effective targeting of EGFR positive GBM cells in the brain. The therapeutic potential of the conjugate was assessed both in vitro, in GBM cell lines and spheroids by the CellTiter-Glo (R) assay, and in vivo using subcutaneous U87-MGvIII xenografts. In addition, mice were imaged pre- and post-PIT using the IVIS/Spectrum/CT to monitor treatment response. Binding of the conjugate correlated to the level of EGFR expression in GBM cell lines. The cell proliferation assay revealed a receptor-dependent response between the tested cell lines. Inhibition of EGFRvIII+ve tumor growth was observed following administration of the immunoconjugate and irradiation. Importantly, this response was not seen in control tumors. In conclusion, the Z(EGFR:03115)-IR700DX showed specific uptake in vitro and enabled imaging of EGFR expression in the orthotopic brain tumor model. Moreover, the proof-of-concept in vivo PIT study demonstrated therapeutic efficacy of the conjugate in subcutaneous glioma xenografts. What's new? Photoimmunotherapy combines the destructive power of photodynamic therapy with the specificity of immunotherapy. But monoclonal antibodies bound to photosensitizers are large, and difficult to get deep into the tumor. Here, the authors attach a photosensitive dye to tiny affibody ligands designed to seek out EGFR, which is often overexpressed in glioblastoma. The affibody molecule brings the dye molecules into the cancer cells, and when it is exposed to near-infrared wavelengths of light. It kills the cell. They showed that the EGFR-affibody combined with the photoreactive dye selectively kills glioblastoma cells overexpressing EGFR in mice.