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High-throughput screening for Survivin and Borealin interaction inhibitors in hepatocellular carcinoma
1State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.
Abstract:
Survivin, a key member of the chromatin passenger complex (CPC), is often highly expressed in human cancers, making it a promising target for cancer treatment. Out of the numerous reported Survivin inhibitors, YM155 is only one entering clinical trial, but was recently failed in the Phase II trial. It is important to develop Survivin inhibitors with new strategies. We recently reported that both Survivin and its binding protein Borealin in the CPC complex are essential for the development of hepatocellular carcinoma, suggesting that disrupting the interaction between Survivin and Borealin would be a promising strategy. Here, we developed a high-throughput screening method based on bimolecular fluorescence complementation (BiFC) technology in cultured cells, which allowed the identification of small chemical inhibitors specifically blocking the Survivin and Borealin interaction. Primary hits from BiFC were further validated in an in vitro AlphaScreen system, which detects the direct interactions of Survivin and Borealin. Etoposide was identified as one of the effective hits. Direct interaction between Survivin and Etoposide was confirmed by surface plasmon resonance assay, and molecular docking analysis suggested the structural information on how Etoposide inhibits the Survivin and Borealin interaction. These results demonstrate a screening system to identify small molecule chemicals inhibiting Survivin and Borealin interaction. In future, an even larger scale screening may lead to identification of better Survivin and Borealin inhibitors.
Insights
Researchers developed a new screening method to find drugs that disrupt the interaction between Survivin and Borealin, crucial proteins in cancer development. Etoposide was identified as a potential inhibitor, offering a new strategy for cancer treatment.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Survivin, a component of the chromatin passenger complex (CPC), is highly expressed in cancers, presenting a therapeutic target.
- Existing Survivin inhibitors have faced clinical setbacks, necessitating novel therapeutic strategies.
- Disrupting the Survivin-Borealin interaction within the CPC is a promising approach for hepatocellular carcinoma treatment.
Purpose of the Study:
- To develop a high-throughput screening (HTS) method to identify small molecules inhibiting the Survivin-Borealin interaction.
- To validate identified inhibitors using in vitro assays and confirm their direct binding to Survivin.
Main Methods:
- Utilized bimolecular fluorescence complementation (BiFC) for high-throughput screening in cultured cells.
- Employed AlphaScreen assays for in vitro validation of Survivin-Borealin interactions.
- Confirmed direct Survivin-Etoposide interaction using surface plasmon resonance (SPR) and molecular docking.
Main Results:
- A BiFC-based screening system successfully identified inhibitors of the Survivin-Borealin interaction.
- Etoposide emerged as a key hit, demonstrating direct binding to Survivin.
- Molecular docking provided insights into the mechanism of Etoposide's inhibition of the Survivin-Borealin interaction.
Conclusions:
- Established a novel screening platform for identifying small molecule inhibitors of the Survivin-Borealin interaction.
- Etoposide shows potential as a therapeutic agent by disrupting this critical protein interaction.
- Further large-scale screening may yield more potent inhibitors for cancer therapy.
