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Dual-Lock System for High Sensitivity and Selectivity in Redox Enzyme Activation and Imaging
So Jin Hong1, Eunsol Jeon2, Min Jung Kim2
1Department of Chemistry, Chung-Ang University, Seoul 06974, South Korea.
Analytical Chemistry
|January 9, 2025
Summary
We developed a novel dual-lock fluorescent probe for detecting cancer hypoxia by simultaneously monitoring two reductases, NTR and hNQO1. This advanced system offers improved in vitro and in vivo hypoxia detection and monitoring capabilities.
Area of Science:
- Biomedical Engineering
- Molecular Imaging
- Chemical Biology
Background:
- Reductase enzyme expression is a key indicator of cellular pathology and is often altered in hypoxic conditions.
- Existing single-detection systems for reductases are limited, leaving a gap in simultaneous dual-reductase detection.
- Cancer hypoxia is a critical factor in tumor progression and treatment resistance, necessitating advanced detection methods.
Purpose of the Study:
- To rationally design and develop a dual-lock fluorescent probe for the simultaneous detection of two specific reductases, NTR and hNQO1.
- To evaluate the probe's performance in detecting hypoxia, particularly in cancer-relevant contexts.
- To demonstrate the probe's utility for noninvasive, real-time monitoring of hypoxia in biological systems.
Main Methods:
- Design of a dual-lock fluorescent probe incorporating a naphthalimide fluorophore with PET and ICT dual-lock control.
- Incorporation of a trimethyl-locked quinone group sensitive to hNQO1 and a nitrobenzyl carbamate group sensitive to NTR.
- Application of the probe in a hypoxia model using HeLa cells and zebrafish embryos for validation.
Main Results:
- The developed dual-lock probe exhibited a high signal-to-noise ratio with an exclusive Off-On fluorescence response to the simultaneous activity of NTR and hNQO1.
- The probe demonstrated superior hypoxia detection in HeLa cells compared to single-detection systems.
- Noninvasive, real-time monitoring of hypoxia was successfully achieved in zebrafish embryos.
Conclusions:
- The dual-lock fluorescent probe provides a novel and effective platform for detecting hypoxia by specifically responding to the concurrent activity of NTR and hNQO1.
- This system offers significant advantages for both in vitro and in vivo applications, enabling advanced diagnostic capabilities.
- The probe facilitates the monitoring of various pathological states characterized by hypoxia.
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