Off-On Squalene Epoxidase-Specific Fluorescent Probe for Fast Imaging in Living Cells.

SQLE (squalene epoxidase) is a cell membrane-bound enzyme. It is a target of fungicides and may become a new target for cancer therapy. Therefore, monitoring the content and distribution of the key enzyme in living cells is very challenging. To achieve this goal, tetraphenyl ethylene-Ter (TPE-Ter) was first designed as a new fluorescent probe to SQLE based on its active cavity. Spectral experiments discovered that SQLE/TPE-Ter shows stronger emission with fast response time and low interference from other analytes. Molecular dynamics simulation clearly confirmed the complex structure of SQLE/TPE-Ter, and the key residues contribute to restriction of TPE-Ter single-molecular motion in the cavity. TPE-Ter-specific response to SQLE is successfully demonstrated in living cells such as LO2, HepG2, and fungi. Imaging of TPE-Ter-treated fungi indicates that it can be used to rapidly assess antifungal drug susceptibility (30 min at least). The present work provides a powerful tool to detect content and distribution of SQLE in living cells.

A simple dual-response fluorescent probe for imaging of viscosity and ONOO- through different fluorescence signals in living cells and zebrafish.

Cellular viscosity is a prominent micro-environmental parameter and peroxynitrite is an essential reactive oxygen special, both of which are involved in various pathological and physiological processes. When the intracellular viscosity is abnormal or the ONOO- concentration is irregular, the normal function of cells will be disturbed. Herein, we rationally designed and synthesized a novel multichannel fluorescent probe (probe 1) for multichannel imaging of viscosity and peroxynitrite. Probe 1 displayed about 108-fold enhancement as the viscosity increased from 1.005 cP to 1090 cP. Moreover, the fluorescence intensity at 540 nm was quickly increased after adding ONOO-. It should be noted that probe 1 has high sensitivity, selectivity and low cytotoxicity, which can be successfully employed for the visualization of exogenous and endogenous ONOO- and imaging viscosity changes in HeLa cells by different fluorescent signals. Furthermore, probe 1 could monitor the change of ONOO- induced by LPS (lipopolysaccharide) and IFN-γ (interferon-γ) in zebrafish. This result reveals that probe 1 may inspire more diagnostic and therapeutic programs for viscosity-peroxynitrite related diseases shortly.

A multifunctional oxygen-producing MnO2-based nanoplatform for tumor microenvironment-activated imaging and combination therapy in vitro.

The current trend of cancer therapy has changed from monotherapy to synergistic or combination therapies. Among the treatment strategies, photodynamic therapy (PDT) and starvation therapy are widely employed together. However, the therapeutic effect of these treatments could lead to strong resistance and poor prognosis due to tumor hypoxia. Therefore, a smart nanoplatform (MONs-GOx@MnO2-Ce6) has been constructed herein by the assembly of glucose oxidase (GOx)-coated mesoporous organosilica nanoparticles (MONs) and MnO2 nanosheets-chlorin e6 (Ce6), which form a nanosystem. Once MONs-GOx@MnO2-Ce6 enter tumor cells, it catalyzes the oxidation of glucose using oxygen (O2) and generates hydrogen peroxide (H2O2) and gluconic acid, the former of which may accelerate the decomposition of MnO2 nanosheets. The released MnO2 nanosheets would regenerate O2 in the presence of H2O2. In this case, MnO2 nanosheets serve as (i) a nanocarrier and fluorescence quencher for the photosensitizer Ce6, (ii) a degradable material that is activated by the tumor microenvironment (TME) for fluorescence recovery, and (iii) an O2-producing carrier that reacts with H2O2 for relieving hypoxia in the tumor, which contributes to the combined starvation/photodynamic cancer therapy since these treatment strategies need O2. MONs-GOx@MnO2-Ce6 could not only realize cancer cell imaging, but also reduce intracellular glucose uptake and Glut1 expression, inhibiting the metabolism of cancer cells. This strategy shows great potential for clinical applications.

Revealing the redox status in endoplasmic reticulum by a selenium fluorescence probe.

As an important organelle, the endoplasmic reticulum (ER) participates in the synthesis and secretion of various proteins, glycogen, lipids and cholesterol in eukaryotic cells. In this work, an endoplasmic reticulum-targeted reversible fluorescent probe (ER-Se) was designed and synthesized. The probe, based on a selenide group, shows high sensitivity and good selectivity toward HClO (LOD = 0.85 µM). In addition, the probe has reversible capability towards HClO/GSH. Most importantly, co-location experiment results indicated that the probe exhibited a great ability to target the endoplasmic reticulum. Furthermore, the probe was successfully applied to detect exogenous and endogenous HClO in ER and monitored the redox status changes during ER stress.

In Vitro Light-Up Visualization of a Subunit-Specific Enzyme by an AIE Probe via Restriction of Single Molecular Motion.

Enzymes contain several subunits to maintain different biological functions. However, it remains a great challenge for specific discrimination of one subunit over another. Toward this end, the fluorescent probe TPEMA is now presented for highly specific detection of the B subunit of cytosolic creatine (CK) kinase isoenzyme (CK-B). Owing to its aggregation-induced emission property, TPEMA shows highly boosted emission toward CK-B with a fast response time and very low interference from other analytes, including the M subunit of CK (CK-M). With the aid of a Job plot assay, ITC assay and molecular dynamics simulation, it was directly confirmed that the remarkably enhanced fluorescence of TPEMA in the presence of CK-B results from the restriction of single molecular motion in the cavity. Selective wash-free fluorescence imaging of CK-B in macrophages under different treatments was successfully demonstrated.

Tumor Microenvironment-Responsive Theranostic Nanoplatform for in Situ Self-Boosting Combined Phototherapy through Intracellular Reassembly.

Through rational design, in vivo supramolecular construction of nanodrugs could precisely proceed in the lesion areas, which may apparently improve the theranostic performance of nanomaterials. Herein, a tumor microenvironment-responsive theranostic nanoplatform (Ce6-GA@MnO2-HA-PEG) has been constructed to achieve in vivo supramolecular construction and enhance the therapeutic efficacy of combined phototherapy through intracellular reassembly. Under the tumor microenvironment, such nanoplatform could undergo the process of decomposition-reassembly and form in situ photothermal assemblies. The generation of assemblies would endow this nanoplatform with the capacity of photothermal therapy. Meanwhile, this nanoplatform could alleviate hypoxia and improve the therapeutic efficacy of photodynamic therapy. The results of in vitro and in vivo experiments reveal that tumors can be ablated efficiently by the designed nanoplatform under laser irradiation. In addition, fluorescence imaging and magnetic resonance imaging can be activated by the decomposition of MnO2 to realize tumor imaging in vivo. Therefore, this multifunctional nanoplatform exhibits the capacity for boosting dual-modal imaging-guided combined phototherapy through intracellular reassembly, which may propose a new thought in cancer theranostics.