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Development of Cell-Based High-Throughput Chemical Screens for Protection Against Cisplatin-Induced Ototoxicity

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Auditory and Vestibular Research

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1427))

Abstract

Various compounds have been tested in recent years for protection against cisplatin-induced hearing loss, but no compound has yet been FDA approved for clinical use in patients. Towards this goal, we developed an unbiased, high-throughput, mammalian cochlear cell-based chemical screen that allowed quantification of the protection ability of bioactive compounds and ranked them for future testing ex vivo in cochlear explant cultures and in vivo in animal models. In our primary screens, protection in the HEI-OC1 organ of Corti immortalized cell line was measured by the ability of each compound to inhibit caspase-3/7 activity triggered by cisplatin treatment (50 μM cisplatin for 22 h). A total of 4385 unique bioactive compounds were tested in a single dose of 8 μM and promising compounds were validated by dose response curves covering ten, 1:3 serial diluted concentrations. Primary hits were defined as having more than 60 % inhibition of the caspase-3/7 activity. Toxicity of the top compounds was measured by a CellTiter-Glo (CTG) assay that measured the viability of the cells in the presence of compound alone in similar dose responsive analysis. A combination of the caspase-3/7 inhibition activity assay (as measured by IC50) and the CTG viability assay (as determined by LD50) identified the top protective compounds in the HEI-OC1 cells. In the future, the top hits in our screens will be tested for their protective ability ex vivo in mouse cochlear explants and in vivo in animal models. Our mammalian cochlear cell-based, high-throughput chemical screening assays described here can be further modified and represent an initial successful step towards therapeutic intervention of hearing disorders, an unmet medical need of our society.

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Acknowledgments

This work was supported in part by grants from the NIH (DC006471, CA21765, DC013879), the Office of Naval Research (N00014-12-1-0191, N00014-12-1-0775, N00014-09-1-1014) and the American Lebanese Syrian-Associated Charities of St. Jude Children’s Research Hospital. We thank Drs. Federico Kalinec and Gilda Kalinec at the House Research Institute for providing the HEI-OC1 cells.

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Authors and Affiliations

  1. Department of Developmental Neurobiology, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN, 38105, USA

    Tal Teitz & Jian Zuo

  2. Department of Chemical Biology and Therapeutics, St. Jude Children’s Research Hospital, 262 Danny Thomas Place, Memphis, TN, 38105, USA

    Asli N. Goktug & Taosheng Chen

Authors
  1. Tal Teitz
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  2. Asli N. Goktug
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  3. Taosheng Chen
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  4. Jian Zuo
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Corresponding author

Correspondence to Jian Zuo .

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Editors and Affiliations

  1. College of Medicine, University of South Florida, Tampa, Florida, USA

    Bernd Sokolowski

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Teitz, T., Goktug, A.N., Chen, T., Zuo, J. (2016). Development of Cell-Based High-Throughput Chemical Screens for Protection Against Cisplatin-Induced Ototoxicity. In: Sokolowski, B. (eds) Auditory and Vestibular Research. Methods in Molecular Biology, vol 1427. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3615-1_22

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  • DOI: https://doi.org/10.1007/978-1-4939-3615-1_22

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3613-7

  • Online ISBN: 978-1-4939-3615-1

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