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A highly selective near-infrared fluorescent probe for imaging H₂Se in living cells and in vivo.

ABSTRACT: Hydrogen selenide (H₂Se), a highly reactive Se species, is an important selenium metabolism intermediate involved in many physiological and pathological processes. This compound is of scientific interest with regard to the real-time monitoring of H₂Se in living cells and in vivo to understand the anti-cancer mechanism of selenium. However, monitoring H₂Se in living cells is still challenging due to the lack of straight forward, highly selective and rapid methods. Here, we developed a novel small-molecule fluorescent probe, NIR-H₂Se, for imaging endogenous H₂Se. NIR-H₂Se exhibited high selectivity toward H₂Se over selenocysteine (Sec), H₂S and small molecule thiols and was successfully used to image the H₂Se content in HepG2 cells during Na₂SeO₃-induced apoptosis. Increased H₂Se content and reduced ROS levels were observed under hypoxic conditions compared to normoxic conditions, which indicated that the cell apoptosis induced by Na₂SeO₃ under a hypoxic environment is via a non-oxidative stress mechanism. Thus, this probe should serve as a powerful tool for exploring the physiological function of H₂Se and Se anticancer mechanisms in a variety of physiological and pathological contexts.

SUBMITTER: Kong F

PROVIDER: S-EPMC5531029 | biostudies-literature | 2016 Feb

REPOSITORIES: biostudies-literature

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A highly selective near-infrared fluorescent probe for imaging H2Se in living cells and in vivo.

Kong Fanpeng F Ge Lihong L Pan Xiaohong X Xu Kehua K Liu Xiaojun X Tang Bo B

Chemical science 20151028 2

Hydrogen selenide (H2Se), a highly reactive Se species, is an important selenium metabolism intermediate involved in many physiological and pathological processes. This compound is of scientific interest with regard to the real-time monitoring of H2Se in living cells and in vivo to understand the anti-cancer mechanism of selenium. However, monitoring H2Se in living cells is still challenging due to the lack of straight forward, highly selective and rapid m ...[more]

PMID: 28808528

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Project description:PurposeCurrent checkpoint inhibitor immunotherapy strategies in glioblastoma are challenged by mechanisms of resistance including an immunosuppressive tumor microenvironment. T cell immunoglobulin domain and mucin domain 3 (TIM3) is a late-phase checkpoint receptor traditionally associated with T cell exhaustion. We apply fluorescent imaging techniques to explore feasibility of in vivo visualization of the immune state in a glioblastoma mouse model.ProceduresTIM3 monoclonal antibody was conjugated to a near-infrared fluorescent dye, IRDye-800CW (800CW). The TIM3 experimental conjugate and isotype control were assessed for specificity with immunofluorescent staining and flow cytometry in murine cell lines (GL261 glioma and RAW264.7 macrophages). C57BL/6 mice with orthotopically implanted GL261 cells were imaged in vivo over 4 days after intravenous TIM3-800CW injection to assess tumor-specific uptake. Cell-specific uptake was then assessed on histologic sections.ResultsThe experimental TIM3-800CW, but not its isotype control, bound to RAW264.7 macrophages in vitro. Specificity to RAW264.7 macrophages and not GL261 tumor cells was quantitatively confirmed with the corresponding clone of TIM3 on flow cytometry. In vivo fluorescence imaging of the 800CW signal was localized to the intracranial tumor and significantly higher for the TIM3-800CW cohort, relative to non-targeting isotype control, immediately after tail vein injection and for up to 48 h after injection. Resected organs of tumor bearing mice showed significantly higher uptake in the liver and spleen. TIM3-800CW was seen to co-stain with CD3 (13%), CD11b (29%), and CD206 (26%).ConclusionsWe propose fluorescent imaging of immune cell imaging as a potential strategy for monitoring and localizing immunologically relevant foci in the setting of brain tumors. Alternative markers and target validation will further clarify the temporal relationship of immunosuppressive effector cells throughout glioma resistance.

Dataset Information

A highly selective near-infrared fluorescent probe for imaging H₂Se in living cells and in vivo.

Publications

A highly selective near-infrared fluorescent probe for imaging H<sub>2</sub>Se in living cells and <i>in vivo</i>.

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