High-throughput screening platform for anticancer therapeutic drug cytotoxicity
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Published on: November 16, 2016
Bhagwant Kaur Sekhon1, Rebecca Heidi Roubin, Aaron Tan
1Faculty of Pharmacy, The University of Sydney, Sydney, New South Wales, Australia.
Abstract:
There are substances that kill cancer cells, but induce T cell proliferation, like thalidomide. To find more of these, a new anticancer drug screening strategy is vital. In this study we report the development of a differential cytotoxicity screening or evaluation platform using the CellTiter-Glo (Promega, Annandale, NSW, Australia) luminescent cell viability assay (ATP assay) and also the CellTiter 96 AQueous (Promega) one solution cell proliferation assay [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay]. The results suggested the platform consisting of the combination of the ATP assay be used for quantifying peripheral blood mononuclear cells, while the more economic MTS colorimetric assay is well suited to be used detecting cell viability of cancer cells. In addition, we found paclitaxel (Taxol, MP Biomedicals Australasia Pty Ltd., Seven Hills, NSW, Australia) to be a useful control for this routine screening methodology. Taxol exhibits the desirable in vitro feature of differential cytotoxicity that spares the immunological cells, when used at a concentration that will kill the majority of the cancer cell population.
Insights
A new drug screening platform identifies anticancer drugs that kill cancer cells while promoting T cell proliferation. This method utilizes ATP and MTS assays for differential cytotoxicity, aiding in the discovery of novel immunomodulatory cancer therapies.
Area of Science:
- Immunology
- Pharmacology
- Biotechnology
Background:
- Thalidomide demonstrates that some anticancer agents can kill cancer cells and stimulate T cell proliferation.
- Identifying similar compounds requires a novel screening strategy to evaluate differential cytotoxicity.
Purpose of the Study:
- To develop and validate a differential cytotoxicity screening platform for identifying anticancer drugs that spare immune cells.
- To assess the utility of ATP and MTS assays in a combined screening approach.
Main Methods:
- Developed a screening platform combining the CellTiter-Glo (ATP) assay for quantifying peripheral blood mononuclear cells (immune cells) and the CellTiter 96 AQueous (MTS) assay for assessing cancer cell viability.
- Utilized paclitaxel (Taxol) as a control drug exhibiting differential cytotoxicity in vitro.
Main Results:
- The ATP assay is suitable for quantifying immune cells, while the MTS assay effectively detects cancer cell viability.
- The combined platform demonstrated differential cytotoxicity, sparing immune cells at concentrations lethal to cancer cells.
- Paclitaxel served as a validated control, showing the desired in vitro characteristics.
Conclusions:
- The developed platform enables the screening of anticancer drugs with differential cytotoxicity, promoting immune cell sparing.
- This strategy is vital for discovering novel anticancer agents that modulate the immune system, like thalidomide.
- The combination of ATP and MTS assays offers an efficient and economic method for routine screening.
