A highly sensitive and selective two-photon fluorescent probe for glutathione S-transferase detection and imaging in...

Area of Science:

• Biochemistry
• Molecular Biology
• Medical Imaging

Background:

• Glutathione transferase (GST) is crucial for metabolizing compounds and is overexpressed in cancer, contributing to drug resistance.
• GST levels are altered in liver injury, making it a key diagnostic biomarker.
• Current methods for monitoring GST activity lack real-time, high-resolution capabilities in living systems.

Purpose of the Study:

• To design and validate a novel fluorescent probe for real-time detection and imaging of glutathione transferase (GST) activity.
• To assess the probe's utility in cancer cells, liver tissues, and in vivo models of liver injury.
• To establish a new tool for studying GST function and associated diseases.

Main Methods:

• Development of a boron-dipyrromethene (BODIPY)-based two-photon probe (BNPA).
• Evaluation of BNPA's selectivity, imaging resolution, and kinetic properties (Michaelis-Menten) towards GSTs.
• Application of BNPA for fluorescence imaging in cancer cells and liver tissues using two-photon microscopy.
• Monitoring of GST activity changes in an $\alpha$-naphthylisothiocyanate (ANIT)-induced liver injury model.

Main Results:

• BNPA demonstrated high selectivity and ultrahigh imaging resolution for GST detection.
• The probe exhibited classic Michaelis-Menten kinetics with GST enzymes.
• BNPA successfully visualized GST activity in living cells and deep tissues via two-photon imaging.
• The probe detected decreased GST activity during ANIT-induced liver injury, correlating with disease progression.

Conclusions:

• BNPA is a promising tool for evaluating GST function and cellular processes in living systems.
• The probe provides a novel approach for studying GST-associated liver diseases.
• BNPA has potential applications in rational drug use and clinical disease diagnosis.