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

Abstract

Rapid and accurate intraoperative pathological diagnosis (IOPD) is essential for intraoperative decision-making to improve patients' outcomes and avoid reoperations. In this study, using a NAD(P)H-activated fluorescent probe, a multifunctional fluorescent indicator has been developed to selectively identify tumor cells from normal tissue and to achieve cancer grading identification. This rapid response probe, CyQ-1, features unprecedented sensitivity and rapid response toward NADH at low nanomolar levels under physiological conditions. Moreover, this indicator allows both colorimetric and fluorescent NADH detection in HeLa, A549, MDA-MB-231, 4T1, MCF-7, HePG2, HUVEC, and HL-7702 cells. Expanding the use of this indicator to advanced tissue models, its ability to visualize NADH in 120 paraffin-embedded colorectal sections and 20 cases of intraoperative frozen sections of lung cancer was further verified. CyQ-1-based cancer grading identification shows an overall 92.5 and 100% agreement with the "gold standard test" of histologic grading toward paraffin and frozen sections, respectively. The sensitivity and specificity for discriminating poorly, moderately, and well-differentiated tumor sections were all above 90%. In a word, the rapid and accurate NADH detection ability for clinical sections makes this proposed indicator a potential candidate for clinical IOPD quantification and tumor differentiation grade recognition.

Near-Infrared Fluorescent Probe with pH- and Viscosity-Switchable Performance for the Detection of Thrombi in Live Animals and Organs

“…Carrageenan was used to induce blood viscosity change and produce blood clots in live mice. The mice had black tails due to blood clots. − The exogenous viscosity change was measured in mice via QL2 . After exogenous injection of 50%, 70%, 80%, and 90% glycerol, the fluorescence of the blood clot group in both channels gradually increased, compared to that of the control group (Figure ).…”

Non-invasive diagnosis of bacterial and non-bacterial inflammations using a dual-enzyme-responsive fluorescent indicator

“…In this study, heterocycle-quinolinium and 5-nitro-indole were selected as specific recognition groups for NAD(P)H and NTR, respectively, based on our previous work. 29,36 These recognition groups were linked to corresponding fluorophores through a polymethine chain to form the indicator CyQ (Fig. 1A).…”

“…The 3-position substituted quinoline salt is well-known for its inherent tendency to undergo two-electron reduction reactions with NAD(P)H analogs. 27,29 Upon the NAD(P)H-mediated reaction, a hydride is transferred from NAD(P)H to the acceptor moiety of quinolinium, resulting in the formation of a novel acceptor–π–donor (A–A–π–D) system exhibiting cyanine characteristics (Fig. 1A).…”

“…A group of scientists has pioneered a range of small-molecule fluorescent probes specifically designed for the individual detection of NAD(P)H and NTR, as exemplified by the work of König, Bhuniya, Tang, Yin, Duan, Chan, Lin, Zhang, Wu, Hu, etc. 23–34 Although promising, until now, the field of concurrent dual-enzyme detection using a fluorescent probe has remained unexplored. In this study, we embarked upon an investigation leveraging a synthetically engineered dual-responsive probe to elucidate variations in NAD(P)H levels across HUVEC cells stimulated by varying concentrations of LPS.…”

A NIR Dual-Channel Fluorescent Probe for Fluctuations of Intracellular Polarity and H2O2 and Its Applications for the Visualization of Inflammation and Ferroptosis

“…According to the relevant literature, one of the most common mediators of inflammation leading to acute liver injury has been identified as lipopolysaccharides (LPSs) . LPS concentrations correlate with the degree of hepatic dysfunction, emphasizing that the progression of hepatic failure is a driver of endotoxemia in patients with chronic liver disease. − Similarly, ferroptosis can elicit specific responses and contribute to the progression of liver disease, as evidenced by the subsequent development of inflammation and eventual fibrosis, cirrhosis, or liver failure. Ferroptosis is a form of cell death mediated by reactive oxygen species (ROS) that is distinct from apoptotic forms of cell death, , a process induced by iron accumulation, reactive oxygen species (ROS) production, and subsequent lipid hydroperoxide. , Characterization of ferroptosis involves the production of soluble and lipid reactive oxygen species (ROS) via iron-dependent heme oxygenase. , There is evidence that elevated hydrogen peroxide levels and fluctuations in intracellular polarity have been found to be associated with human diseases, including inflammatory diseases and ferroptosis. , Excessive H 2 O 2 causes hepatocytes to produce high levels of inflammatory factors while inducing lipid peroxidation, leading to ferroptosis.…”