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Basis of a FTIR spectroscopy methodology for automated evaluation of Akt kinase inhibitor on leukemic cell lines used as model

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Abstract

The PI3K/Akt-signaling pathway, associated with cancer development and disease progression, is recognized to be an anti-tumor drug target that could present important therapeutic benefit. However, no targeted Akt medicines have been commercialized yet, reflecting that drug selection procedures requires significant improvement from early research to clinical trials. Thus, new methods permitting both the evaluation of cytotoxic and proliferation inhibition effect on cancer cells but also to provide a global fingerprint of the drug action mechanism of new Akt inhibitor candidates are of major interest. Because it can detect very subtle molecular changes and could provide a global fingerprint of drug effects on cells, Fourier-transform infrared (FTIR) spectroscopy appears to be a promising method to develop new time- and cost-saving tools for chemical library screening improvements. In this study, we combine FTIR spectroscopy, advanced chemometrics analysis and cross-validation by standard biological assays to establish a basis of a mid-throughput methodology for rapid and automated assessment of cell response to Akt inhibitors and quantitative evaluation of their anti-proliferative effects. Our results shows that our methodology is able (1) to detect cell response to an Akt inhibitor exposure even for very low doses, (2) to provide biochemical information of interest about its effects on the cell metabolism, lipidome, and proteome, (3) to predict accurately resulting cell proliferation inhibition rate. Thus, further based on a large spectral data base, our methodology could contribute to facilitate preliminary screening of chemical libraries and improving the selection procedure of drug candidates in laboratory routine.

Validation performance in prediction with PLS regression of the cell proliferation inhibition induced by A6730 on K562 cells. Blue average proliferation inhibition (±std) as assessed by MTS on five experiments; Red average proliferation inhibition predicted by PLS regression on three independent experiments; R 2 correlation coefficient; RMSEV root mean square error of validation

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Acknowledgments

The authors thank the FFR “Fédération Française de Rugby” for its technical contribution through the loan of a HTS-XT device coupled to a Tensor-27 FTIR spectrometer (Brüker Optics, Ettlingen, Germany) within the framework of the research on the cancer. The authors would like also to thanks Dr. Dominique Bertrand, Mr. Teddy Hapillon, Dr. Cyril Gobinet and Dr. Benjamin Bird for helpful MATLAB programming advices and tips.

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Authors and Affiliations

  1. University Bordeaux, Pharmacochimie, FRE 3396, 33000, Bordeaux, France

    Adrian Travo, Vanessa Desplat, Emmanuelle Barron, Emilie Poychicot-Coustau, Jean Guillon, Gérard Déléris & Isabelle Forfar

  2. CNRS, Pharmacochimie, FRE 3396, 33000, Bordeaux, France

    Adrian Travo, Vanessa Desplat, Emmanuelle Barron, Emilie Poychicot-Coustau, Jean Guillon, Gérard Déléris & Isabelle Forfar

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  1. Adrian Travo
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  2. Vanessa Desplat
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  3. Emmanuelle Barron
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  4. Emilie Poychicot-Coustau
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  5. Jean Guillon
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  6. Gérard Déléris
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  7. Isabelle Forfar
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Corresponding author

Correspondence to Isabelle Forfar.

Additional information

Gérard Déléris passed away on January 19, 2012.

This paper is dedicated to the memory of Prof. Gérard Déléris.

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Travo, A., Desplat, V., Barron, E. et al. Basis of a FTIR spectroscopy methodology for automated evaluation of Akt kinase inhibitor on leukemic cell lines used as model. Anal Bioanal Chem 404, 1733–1743 (2012). https://doi.org/10.1007/s00216-012-6283-1

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  • DOI: https://doi.org/10.1007/s00216-012-6283-1

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