Fluorescence and ratiometric photoacoustic imaging of endogenous furin activity via peptide functionalized MoS2 nanosheets

Abstract

Furin is an important cellular endoprotease, which is expressed at high levels in various cancer cells. Accurate and real-time detection of endogenous furin with high sensitivity and selectivity is significant for the diagnosis of cancer. Herein an activatable nanoprobe (MoS₂@PDA–PEG/peptide, MPPF) with dual-mode near-infrared fluorescence (NIRF)/ratiometric photoacoustic (PA) imaging of endogenous furin activity has been developed. The MPPF nanoprobes were constructed by the covalent functionalization of polydopamine (PDA) coated MoS₂ nanosheets (NSs) with Cy7-labeled furin substrate peptides. Upon cleavage of the peptides by furin, Cy7 molecules are released from MPPF nanoprobes and recover their fluorescence, realizing furin activity detection with the limit of detection (LOD) down to 3.73 × 10⁻⁴ U mL⁻¹. Meanwhile, the ratio of the PA signal at 768 nm to that at 900 nm (PA₇₆₈/PA₉₀₀) decreases over time due to the destruction of fluorescence resonance energy transfer effect from Cy7 to MoS₂ NSs and the rapid clearance of small Cy7 molecules from tissues. Thus, the simultaneous change in NIRF and ratiometric PA signals enables the imaging of endogenous furin activity in real time, and with high sensitivity, and high selectivity in both tumor cells and tumor-bearing mice.