Demystifying Lysosomal α-l-Fucosidase in Liver Cancer-Bearing Mice by Specific Two-Photon Fluorescence Imaging

Liver cancer is one of the most frequently diagnosed cancers and has high mortality. However, the early treatment and prognosis can greatly prolong the survival time of patients, which depends on its early detection. α-l-Fucosidase (AFU), as a vital lysosomal hydrolase, is considered to be an ideal biomarker for early stage liver cancer. So, in vivo monitoring of AFU is essential for the early and accurate diagnosis of liver cancer. Hence, we designed the first two-photon turn-on fluorescent reporter, termed HcyCl-F, which localized to lysosomes for fast imaging of AFU. The 2-chloro-4-phenyl-α-l-fucoside bond of HcyCl-F could be effectively hydrolyzed by AFU and released the hydroxyl on the benzene ring, eventually obtaining a strong conjugated compound (HcyCl-OH) with shiny fluorescence. We demonstrated that HcyCl-F was able to rapidly and accurately respond to AFU. Using a two-photon fluorescence microscope, we successfully visualized the fluctuation of AFU in lysosomes. More importantly, a fascinatingly strong fluorescence signal was observed in the tumor tissue of liver cancer-bearing mice. Of note, we confirmed that HcyCl-F could clearly detect liver tumors in stage I. Altogether, our work provides a simple and convenient method for deciphering the critical pathological function of AFU in depth and facilitates the nondestructive and effective diagnosis of liver cancer in the early stage.