Bright and Specific Targeting of Metastatic Lymph Nodes in Orthotopic Mouse Models of Gastric Cancer with a Fluorescent Anti-CEA Antibody.

BACKGROUND

Gastric cancer poses a major diagnostic and therapeutic challenge. Improved visualization of tumor margins and lymph node metastases with tumor-specific fluorescent markers could improve outcomes.

METHODS

To establish orthotopic models of gastric cancer, one million cells of the human gastric cancer cell line, MKN45, were suspended in 50 μl of equal parts PBS and Matrigel and injected into the nude mouse stomach with a 29-gauge needle. Tumors were allowed to grow for 8-12 weeks before performing imaging studies. For tumor labeling, M5A (humanized anti-CEA mAb) and IgG as a control, were conjugated with the near-infrared dye IRDye800CW. Mice were randomized to receive 50 μg of M5A-IR800 (n = 14) or 50 μg of IgG-IR800 (n = 14) intravenously and were imaged 72 hours later. Fluorescence imaging was performed using the LI-COR Pearl Imaging System.

RESULTS

Forty-two lymph nodes were collected from 28 mice, of which 59.5% were found to contain metastatic gastric cancer cells on pathologic examination. In mice that received M5A-IR800, there was a statistically significant difference in the mean fluorescence signal for cancer-positive lymph nodes at 0.431 (SE ± 0.224) compared with 0.105 (SE ± 0.009) for cancer-negative nodes (p: 0.002). For IgG-IR800, there was no significant difference in the mean fluorescence signal for cancer-positive nodes (0.057) compared with cancer-negative nodes (0.064), p-value 0.677.

CONCLUSIONS

Humanized anti-CEA (M5A) antibodies conjugated to fluorescent dyes provide bright labeling of lymph nodes containing metastatic gastric cancer. This tumor-specific fluorescent antibody is a promising clinical tool for identifying lymph nodes containing metastatic gastric cancer.