An Activatable NIR Fluorescent Probe for NAD(P)H and Its Application to the Real‐Time Monitoring of p53 Abnormalities In Vivo

Abstract

NAD(P)H is crucial for biosynthetic reactions and antioxidant functions. However, the current probes developed for detecting NAD(P)H in vivo require intratumoral injection, which limited their application for animal imaging. To address this issue, we have developed a liposoluble cationic probe, KC8, which exhibits excellent tumor‐targeting ability and near‐infrared (NIR) fluorescence after reaction with NAD(P)H. By using KC8, it was demonstrated for the first time that the level of NAD(P)H in the mitochondria of living colorectal cancer (CRC) cells was highly related to the abnormality of the p53. Furthermore, KC8 was successfully used to differentiate not only between tumor and normal tissue but also between tumors with p53 abnormality and normal tumors when administered intravenously. Finally, we evaluated tumor heterogeneity through two fluorescent channels after treating a tumor with 5‐Fu. This study provides a new tool for real‐time monitoring of the p53 abnormality of CRC cells.

NAD(P)H Activated Fluorescent Probe for Rapid Intraoperative Pathological Diagnosis and Tumor Histological Grading

“…The NMR spectra were recorded at room temperature with a Bruker Vance III HD 400 MHz spectrometer. The 1 H NMR spectra were recorded at 400 MHz, and the 13 C NMR spectra at 100 MHz. The purified water used in all experiments was produced by a Milli-Q system (Millipore, >18.0 MΩ, USA).…”

“…NAD(P)H indicators (CyQ-1 and CyQ-2) and the reaction intermediates were synthesized conveniently without the requirement of chromatography purification steps (Figure S1). The final products were characterized by 1 H NMR, 13 C NMR, mass spectrometry, and UV−vis spectroscopy, as shown in Figures S2−S9.…”

“…11 NAD(P)H has long been used as an important intracellular bioprobe to noninvasively monitor and analyzes the metabolic activity of living cells and tissues. 12,13 Cancer cells are reported to have a high rate of NAD(P) + / NAD(P)H turnover due to their elevated metabolic needs. 14,15 Increased NAD(P)H levels enhance glycolysis and fuel cancer cells, 16 which makes NAD(P)H biosensors potential candidates for distinguishing a tumor from normal tissue and determining tumor differentiation grade.…”

“…1,4-Dihydronicotinamide adenine dinucleotide (NADH) and its phosphate ester (NADPH) are key carriers of electrons in cellular energy metabolism and the indispensable redox regulator in live cells . NAD­(P)H has long been used as an important intracellular bioprobe to noninvasively monitor and analyzes the metabolic activity of living cells and tissues. , Cancer cells are reported to have a high rate of NAD­(P) + /NAD­(P)H turnover due to their elevated metabolic needs. , Increased NAD­(P)H levels enhance glycolysis and fuel cancer cells, which makes NAD­(P)H biosensors potential candidates for distinguishing a tumor from normal tissue and determining tumor differentiation grade. However, the lack of efficient probes has restrained the investigation of NAD­(P)­H-dependent pathways in clinically rapid pathological diagnosis.…”

Fluorescent dyes based on rhodamine derivatives for bioimaging and therapeutics: recent progress, challenges, and prospects

“…48). 324 Preliminary evaluation revealed the increase in the efficiency of KC8 to promote the specific reaction of NADH with quinoline moieties after the introduction of borate ester groups. After treatment with NAD(P)H in PBS, the fluorescence intensity of KC8 at 745 nm increased by more than 6-fold.…”