Rapid and Accurate Visualization of Breast Tumors with a Fluorescent Probe Targeting -Mannosidase 2C1

Accurate detection of breast tumors and discrimination of tumor from normal tissues during breast-conserving surgery are essential to reduce the risk of misdiagnosis or recurrence. However, existing probes show substantial background signals in normal breast tissues. In this study, we focus on glycosidase activities in breast tumors. We synthesized a series of 12 fluorescent probes and performed imaging-based evaluation on surgically resected human breast specimens. Among them, the alpha-mannosidase-reactive fluorescent probe HMRef-alpha Man detected breast cancer with 90% sensitivity and 100% specificity. We identified alpha-mannosidase 2C1 as the target enzyme and confirmed its overexpression in various breast tumors. We found that fibroadenoma, the most common benign breast lesion in young woman, tends to have higher alpha-mannosidase 2C1 activity than malignant cancer. Combined application of green-emitting HMRef-alpha Man and a red-emitting gamma-glutamyltranspeptidase probe enabled efficient dual-color, dual-target optical discrimination of malignant and benign tumors.