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Screening compound libraries for H2O2-mediated cancer therapeutics using a peroxiredoxin-based sensor
Yining Hao1, Troy F Langford1, Sun Jin Moon1
1Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Abstract:
Compounds that modulate H2O2 reaction networks have applications as targeted cancer therapeutics, as a subset of cancers exhibit sensitivity to this redox signal. Previous studies to identify therapeutics that induce oxidants have relied upon probes that respond to many different oxidants in cells, and thus do not report on only H2O2, a redox signal that selectively oxidizes proteins. Here we use a genetically encoded fluorescent probe for human peroxiredoxin-2 (Prx2) oxidation in screens for small-molecule compounds that modulate H2O2 pathways. We further characterize cellular responses to several compounds selected from the screen. Our results reveal that some, but not all, of the compounds enact H2O2-mediated toxicity in cells. Among them, SMER3, an antifungal, has not been reported as an oxidant-inducing drug. Several drugs, including cisplatin, that previously have been shown to induce reactive oxygen species (ROS) do not appear to oxidize Prx2, suggesting H2O2 is not among the ROS induced by those drugs.
Insights
Researchers screened compounds targeting hydrogen peroxide (H₂O₂) pathways using a novel fluorescent probe. Some compounds, like the antifungal SMER3, showed H₂O₂-mediated toxicity, while others, including cisplatin, did not target this specific redox signal.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Therapeutics
Background:
- Hydrogen peroxide (H₂O₂) is a key redox signal with therapeutic potential in cancer.
- Existing methods for identifying H₂O₂-modulating compounds lack specificity.
- Cancers sensitive to H₂O₂ offer a target for novel therapeutics.
Purpose of the Study:
- To identify small-molecule compounds that specifically modulate H₂O₂ pathways.
- To characterize cellular responses to identified H₂O₂-modulating compounds.
- To investigate the role of H₂O₂ in drug-induced oxidative stress.
Main Methods:
- Utilized a genetically encoded fluorescent probe for human peroxiredoxin-2 (Prx2) oxidation.
- Conducted high-throughput screening of small-molecule libraries.
- Performed cellular assays to assess compound-induced H₂O₂-mediated toxicity.
- Differentiated H₂O₂-specific effects from general reactive oxygen species (ROS) induction.
Main Results:
- Identified several small-molecule compounds that modulate H₂O₂ pathways.
- Demonstrated that some compounds induce H₂O₂-mediated cellular toxicity.
- Revealed that the antifungal SMER3 acts as an oxidant-inducing drug.
- Found that established ROS-inducing drugs like cisplatin do not oxidize Prx2, indicating they do not primarily induce H₂O₂.
Conclusions:
- A novel fluorescent probe enables specific screening for H₂O₂ pathway modulators.
- SMER3 is a potential H₂O₂-targeting therapeutic agent.
- Drug-induced ROS are not always mediated by H₂O₂, highlighting the need for specific probes.
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