Proximity-Induced Hybridization Chain Reaction-Based Photoacoustic Imaging System for Amplified Visualization Protein-Specific Glycosylation in Mice

Glycosylation is a key cellular mechanism that regulates several physiological and pathological functions. Therefore, identification and characterization of specific-protein glycosylation in vivo are highly desirable for studying glycosylation-related pathology and developing personalized theranostic modalities. Herein, we demonstrated a photoacoustic (PA) nanoprobe based on the proximity-induced hybridization chain reaction (HCR) for amplified visual detection of protein-specific glycosylation in vivo. Two kinds of functional DNA probes were designed. A glycan probe (DBCO-GP) was attached to glycans through metabolic oligosaccharide engineering (MOE) and protein probe (PP)-targeted proteins by aptamer recognition. Proximity-induced hybridization of the complementary domain between the two kinds of probes promoted conformational changes in the protein probes and in situ release of the HCR initiator domain. Gold nanoparticles (AuNPs) modified by complementary sequences (Au-H1 and Au-H2) self-assembled into Au aggregates via the HCR, thereby converting DNA signals to photoacoustic signals. Due to the high contrast and deep penetration of photoacoustic imaging, this strategy enabled in situ detection of Mucin 1 (MUC1)-specific glycosylation in mice with breast cancer and successfully monitored its dynamic states during tunicamycin treatment. This imaging technique provides a powerful platform for studying the effects of glycosylation on the protein structure and function, which helps to elucidate its role in disease processes.