Knock-in reporter constructs and drug screening for compounds to restore RKIP expression in cancer
Sawssen Bouali1, Xunzhen Zheng2, Chris Figy2
1Department of Medical Chemistry, University of Szeged, 6720, Szeged, Dóm tér 8, Hungary.
Abstract:
RKIP (Raf Kinaes Inhibitory Protein) is ubiquitously expressed in almost all normal tissues of metazoans. Consistent with its negative regulatory role in cell proliferation and survival, RKIP expression is progressively downregulated in cancer accompanied with a worse prognosis and disease outcome. Experiments with cancer cell lines and genetically engineered mice have demonstrated that the expression level of RKIP is a driving factor in determining the disease outcome of cancer, and that restoring RKIP expression is a promising option for therapeutic treatment of low-RKIP-expressing cancers. RKIP expression is mainly regulated at the transcription level such that its expression is tuned down, but not shut off, and is poised to be reactivated. In this study we developed a RKIP gene promoter knock-in reporter breast cancer cell line to screen for compounds that can regulate RKIP transcription in cancer. Using this modified cell line, we were able to identify eight FDA approved compounds that increase RKIP promoter activity by a minimum of 2-fold and can potentially be re-purposed for therapeutic treatment of low-RKIP-expressing breast cancer.
Insights
Raf Kinase Inhibitory Protein (RKIP) is often lost in cancer, worsening prognosis. This study identified eight FDA-approved drugs that can restore RKIP expression, offering potential new breast cancer therapies.
Area of Science:
- Molecular Biology
- Cancer Research
- Pharmacology
Background:
- Raf Kinase Inhibitory Protein (RKIP) is crucial for regulating cell proliferation and survival.
- RKIP expression is significantly downregulated in various cancers, correlating with poor prognosis.
- Restoring RKIP levels presents a promising therapeutic strategy for cancers with low RKIP expression.
Purpose of the Study:
- To develop a reporter system for screening compounds that modulate RKIP transcription in breast cancer.
- To identify existing FDA-approved drugs that can upregulate RKIP expression for potential repurposing.
Main Methods:
- Creation of a breast cancer cell line with a knock-in reporter for the RKIP gene promoter.
- High-throughput screening of FDA-approved compounds using the developed reporter cell line.
- Quantification of RKIP promoter activity to identify compounds that increase transcription.
Main Results:
- Eight FDA-approved compounds were identified that significantly increase RKIP promoter activity (minimum 2-fold).
- These compounds demonstrate the potential to restore RKIP expression in breast cancer cells.
- The developed reporter cell line is effective for screening transcriptional regulators of RKIP.
Conclusions:
- RKIP transcription is a viable target for therapeutic intervention in breast cancer.
- Repurposing FDA-approved drugs offers a rapid path for developing new treatments for low-RKIP-expressing breast cancers.
- This study provides a foundation for further investigation into RKIP-based cancer therapies.
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