Multifunctional Fluorescent Probe for Simultaneous Detection of ONOO^–, Viscosity, and Polarity and Its Application in Ferroptosis and Cancer Models

Abstract

Intracellular peroxynitrite anions (ONOO–) and microenvironments (such as viscosity and polarity) play an important role in maintaining redox homeostasis, regulating diffusion, transportation, and signal transduction in living cells. The abnormality of these factors is often closely related to various physiological/pathological processes. However, owing to the lack of suitable probes, the simultaneous visualization of ONOO–, viscosity, and polarity in ferroptosis and cancer models has not been achieved. To meet urgent needs, we presented a multifunctional near-infrared (NIR) fluorescent probe, named MQA-P, for simultaneously detecting ONOO–, viscosity, and polarity within mitochondria. The probe exhibited a remarkable turn-on response to ONOO– with the far-red emission of about 645 nm and was highly sensitive to viscosity/polarity in the NIR channel with λem > 704 nm. Facilitated by MQA-P, for the first time, we revealed that erastin-induced ferroptosis was accompanied by a significant upregulation of ONOO– and an increase of viscosity (or decrease of polarity) at the same time. Moreover, the concurrent use of ONOO–, viscosity, and polarity for the diagnosis of cancer has been successfully achieved not only at cell/tissue levels but also in tumor mice models. Compared with detecting only one factor, this simultaneous detection of multimarkers provides a more sensitive and reliable method/tool for tracking ferroptosis-related pathological processes and cancer diagnosis, holding great potential in preclinical research, medical diagnosis, and imaging-guided surgery.

A wash-free fluorescent probe with a large Stokes shift for the identification of NAFL through tracing the change of lipid droplets

“…Fluorescent probes have become a popular tool for real-time imaging and exploration of various physiological processes due to their advantages of high sensitivity, excellent resolution, non-invasiveness, and fast response compared to other technologies. 11 To date, a large number of fluorescent probes have been developed for the monitoring of various physiological processes in LDs. 12–14 For instance, in 2022, the team of Peiyao Xie 15 designed a fluorescent probe TPA-SO2 , which can simultaneously detect SO 2 and LD polarity during ferroptosis.…”

Dual Ratio and Ultraprecision Quantification of Mitochondrial Viscosity in Ferroptosis Enabled by a Vibration-Based Triple-Emission Fluorescent Probe

“…These functional and structural changes forebode the mitochondrial abnormality. − Viscosity, as one of the crucial microenvironmental parameters for characterizing the mitochondrial state, − is believed that monitoring its changes will help us to understand the function during ferroptosis. By monitoring the changes in fluorescence intensity, Dong and co-workers discovered that the increased viscosity occurred during the ferroptosis . However, since intensity-dependent probes are easily disturbed by various factors (external microenvironments, instrument parameters, photobleaching), they are not appropriate for accurate analysis. − Therefore, there is an urgent need to develop a precise tool for the quantitative analysis of mitochondrial viscosity changes during ferroptosis.…”

“…By monitoring the changes in fluorescence intensity, Dong and co-workers discovered that the increased viscosity occurred during the ferroptosis. 18 However, since intensity-dependent probes are easily disturbed by various factors (external microenvironments, instrument parameters, photobleaching), they are not appropriate for accurate analysis. 19−21 Therefore, there is an urgent need to develop a precise tool for the quantitative analysis of mitochondrial viscosity changes during ferroptosis.…”

ONOO– Activatable Fluorescent Sulfur Dioxide Donor for a More Accurate Assessment of Cell Ferroptosis

“…•− ) reacts with nitric oxide (NO). 40 It has previously been reported that ONOO − can promote oxidative damage and induce lipid peroxidation, and it can further trigger ferroptosis. 41 The ROS changes will therefore be appropriately reflected by reliable and precise ONOO − detection, which may significantly aid in the elucidation of its biological consequences during ferroptosis and further enable thorough investigation and manipulation of ferroptosis.…”

“…Peroxynitrite (ONOO – ) is a type of ROS that is usually generated when superoxide (O 2 •– ) reacts with nitric oxide (NO) . It has previously been reported that ONOO – can promote oxidative damage and induce lipid peroxidation, and it can further trigger ferroptosis .…”