NIR-II Navigation with an EGFR-Targeted Probe Improves Imaging Resolution and Sensitivity of Detecting Micrometastases in Esophageal Squamous Cell Carcinoma Xenograft Models

Abstract

The survival rate of esophageal squamous carcinoma (ESCC) after surgical resection is estimated to be only 30.3% due to the difficulty in identifying microinfiltration and subtle metastases. In this study, we explored the value of near-infrared fluorescence in the second window (NIR-II) using an epidermal growth factor receptor (EGFR)-targeted probe (cetuximab-IR800) for the intraoperative navigation of ESCC in xenograft mouse models. Immunohistochemical results showed that EGFR was aberrantly expressed in 94.49% (120/127) of ESCC tissues and 90.63% (58/64) of metastatic lymph nodes. Western blot results demonstrated that EGFR protein was highly expressed in ESCC cell lines. Flow cytometry data revealed that cetuximab-IR800 showed a stronger binding specificity in EGFR-positive KYSE-30 cells than in A2780 control cells (P < 0.01). In vivo imaging data showed that the ratio of mean fluorescent intensity (MFI) and tumor to background (TBR) was significantly higher in KYSE-30 subcutaneous tumors with the infusion of cetuximab-IR800 than in those with the infusion of IgG1-IR800 (P < 0.05). Surgical navigation with NIR-II imaging showed that the TBR in orthotopic ESCC was significantly higher than that of NIR in the first window (NIR-I) (2.11 ± 0.46 vs 1.58 ± 0.31, P < 0.05), and NIR-II was more sensitive than NIR-I in detecting subcentimeter metastases (94.87% (37/39) vs 58.97% (23/39), P < 0.001). In conclusion, cetuximab-IR800 with high specificity for ESCC was first used in NIR-II surgical navigation. This probe showed better imaging resolution and higher sensitivity in detecting subtle metastases derived from an orthotopic ESCC model than NIR-I, which indicates that NIR-II has promise in guiding precise surgery for ESCC patients.

Near-infrared dye IRDye800CW-NHS coupled to Trastuzumab for near-infrared II fluorescence imaging in tumor xenograft models of HER-2-positive breast cancer

“…The molar extinction coefficients express by ε , ε dye and ε antibody are 240 000 and 203 000 L mol −1 cm −1 , respectively. 24,30 The dye-antibody ratio for Trastuzumab-800CW is calculated to be 2.44 : 1.…”

“…The molar extinction coefficients express by e, e dye and e antibody are 240 000 and 203 000 L mol À1 cm À1 , respectively. 24,30 The dyeantibody ratio for Trastuzumab-800CW is calculated to be 2.44 : 1. Then, we investigated the fluorescence properties of IRDye800-CW-NHS, ICG-NHS, Trastuzumab-800CW and Trastuzumab-ICG.…”

“…22,23 The recently reported IRDye800CW is both biocompatible and easy to modify, the modified IRDye800CW can be directly reacted with biomolecules. [24][25][26] And the IRDye800CW is being evaluated for reactive modifications in multiple clinical trials. 27 However, the effect of fluorophores on targeting performance is significant, and no systematic comparative study of the performance of the two fluorescent dyes and their binding to targeting molecules for imaging in the NIR-II has been performed.…”

Identification of an IGF2BP2-Targeted Peptide for Near-Infrared Imaging of Esophageal Squamous Cell Carcinoma

“…Given the better tissue penetration and tumor signal-to-noise ratio, NIR in the second window (NIR-II) tumor-targeted imaging obviously holds greater potential for clinical translation [ 32 , 33 ]. Imaging of deep tumors with NIR-II dyes such as IRDye800cw will undoubtedly become more widespread [ 5 , 10 ]. In addition, short peptides have demonstrated their superior performance in tumor diagnostic applications [ 11 , 24 ].…”

“…Novel imaging systems have emerged in response to the limited effectiveness of conventional white light endoscopy in the routine diagnoses of ESCC. Due to the remarkably low absorption and autofluorescence of biological tissues within the near-infrared (NIR) spectrum (650–900 nm), near-infrared fluorescence (NIRF) imaging has been employed to visualize various biomolecules which are closely related to the disease’s processes at the cellular and even molecular levels [ 5 , 6 ].…”