Intraoperative fluorescence delineation of head and neck cancer with a fluorescent Anti‐epidermal growth factor receptor nanobody

Abstract

Intraoperative near-infrared (NIR) fluorescence imaging is a technology with high potential to provide the surgeon with real-time visualization of tumors during surgery. This study explores the feasibility for clinical translation of an epidermal growth factor receptor (EGFR) targeting nanobody for intraoperative imaging and resection of orthotopic tongue tumors and cervical lymph node metastases. The anti-EGFR nanobody 7D12 and the negative control nanobody R2 were conjugated to the NIR fluorophore IRDye800CW (7D12-800CW and R2-800CW). Orthotopic tongue tumors were induced in nude mice using the OSC-19-luc2-cGFP cell line. Tumor bearing mice were injected with 25µg 7D12-800CW, R2-800CW or 11µg 800CW. Subsequently, other mice were injected with 50 µg or 75 µg of 7D12-800CW. The FLARE imaging system and the IVIS spectrum were used to identify, delineate and resect the primary tumor and cervical lymph node metastases. All tumors could be clearly identified using 7D12-800CW. A significantly higher tumor-to-background ratio (TBR) was observed in mice injected with 7D12-800CW compared to mice injected with R2-800CW and 800CW. The highest average TBR (2.00 ± 0.34 and 2.72 ± 0.17 for FLARE and IVIS spectrum, respectively) was observed 24 hours after administration of the EGFR-specific nanobody. After injection of 75 µg 7D12-800CW cervical lymph node metastases could be clearly detected. Orthotopic tongue tumors and cervical lymph node metastases in a mouse model were clearly identified intraoperatively using a recently developed fluorescent EGFR targeting nanobody. Translation of this approach to the clinic would potentially improve the rate of radical surgical resections.

Activin Receptor-like Kinase 1 Ligand Trap Reduces Microvascular Density and Improves Chemotherapy Efficiency to Various Solid Tumors

“…KEP1-11 cells with stable expression of Fc, ALK1-Fc, ALK2-Fc, and Flt-Fc were generated by infecting cells with lentiviruses and selection with 5 mg/mL puromycin (Sigma-Aldrich). Luciferaseexpressing human OSC19 head and neck cancer cells were described before (23) and maintained in DMEM-containing 10% FCS, penicillin/streptomycin, MEM-NEAA (1%), MEM vitamin mixture (1%), Glutamax (all Sigma-Aldrich), 1 mmol/L pyruvate (Thermo Scientific).…”

Optimization of fluorescent imaging in the operating room through pulsed acquisition and gating to ambient background cycling

“…Fluorescence cannot only be used to mark sites of pathology for resection but also to mark sensitive areas for the surgeon to avoid [5][6][7][8][9]. Increased availability of specific fluorescent probes which delineate ducts, tissues and/or molecular expression will continue, and these will gradually find successful applications in surgical procedures.…”

TM1-IR680 peptide for assessment of surgical margin and lymph node metastasis in murine orthotopic model of oral cancer

“…Fluorescent-based optical imaging using Near Infrared (NIR) dyes has emerged as an optimal tool for this, as they offer low background and better penetration capability. Optical imaging using NIR fluorophores are feasible for cancers that are not deeply located, like breast cancer, colorectal cancer and oral cancer20. The fluorophores currently used in image-guided surgery for sentinel lymph node mapping and tumour mapping are indocyanine green (ICG) and 5-ALA20.…”