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Computed determination of the in vitro optimal chemocombinations of sphaeropsidin A with chemotherapeutic agents to combat melanomas

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Abstract

Purpose

Evasion to new treatments of advanced melanoma is still associated with a poor prognosis. Choosing the best combination of agents that can bypass resistance mechanisms remains a challenge. Sphaeropsidin A (Sph A) is a fungal bioactive secondary metabolite previously shown to force melanoma cells to undergo apoptosis via cell volume dysregulation. This work studied its in vitro combination with cytotoxic chemotherapeutics in a rational manner.

Methods

Four melanoma cell lines harboring different sensitivity levels to pro-apoptotic stimuli were used to build a predictive response surface model allowing the determination of the optimal in vitro combinations of Sph A with two drugs, i.e., cisplatin or temozolomide, owing to a limited set of experimentations.

Results

Testing 12 experimental combinations allowed us to build an accurate predictive model that considers the complexity of the drug interaction and determines the optimal combinations according to the endpoint chosen, i.e., the maximal cytotoxic effects. Therefore, combining 4 µM Sph A with 75 µM cisplatin concomitantly for 72 h improved its cytotoxic effects on melanoma cells in a synergistic manner. An optimal in vitro treatment schedule was also obtained for temozolomide.

Conclusions

The use of a response surface model offers the possibility of reducing the experiments while determining accurately the optimal combinations. We herein highlighted that combining the Na+/K+/2Cl− cotransporter and/or anion exchanger inhibitor Sph A with chemotherapeutic agents could improve the therapeutic benefits of conventional chemotherapies against advanced melanomas, particularly because Sph A exerts cytotoxic effects regardless of the genetic BRAF and NRAS status.

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Abbreviations

ADME:

Absorption, distribution, metabolism, and excretion

AUC:

Area under the curve

CDDP:

Cisplatin

Cmax:

Maximum (or peak) serum concentration

CTLA-4:

Cytotoxic T lymphocyte antigen-4

DTIC:

Dacarbazine

EDTA:

Ethylenediaminetetraacetic acid

GI50 :

Growth inhibition 50%

GST:

Glutathione-S-transferase

LC50/90 :

Lethal concentration 50/90

MCDM:

Multi-criteria decision-making

MDR:

Multidrug resistance

MTIC:

5-(3-methyltriazen-1-yl)imidazole-4-carboxamide

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

NCI:

National cancer institute

NKCC1/2:

Na+–K+–2Cl− co-transporters ½

PBS:

Phosphate buffered saline

PD1:

Programmed cell death protein 1

PFS:

Progression-free survival

RVI:

Regulatory volume increase

Sph A:

Sphaeropsidin A

TMZ:

Temozolomide

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Acknowledgements

We thank Robert Kiss for his advice in the current project and particularly for having initiated it as well as for reviewing the present manuscript. ADG is a postdoctoral fellow of the FWO-Vlaanderen, Belgium.

Funding

No private or public grant supported this work that was conducted with academic financial support only.

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Authors and Affiliations

  1. Laboratoire de Cancérologie et Toxicologie Expérimentale, Université Libre de Bruxelles, Boulevard du Triomphe, Accès 2, 1050, Ixelles, Belgium

    Aude Ingels & Véronique Mathieu

  2. Department of Medicine I, Institute of Cancer Research, Medical University Vienna, Spitalgasse 23, 1090, Vienna, Austria

    Carina Dinhof & Walter Berger

  3. Comprehensive Cancer Center, Medical University Vienna, Spitalgasse 23, 1090, Vienna, Austria

    Carina Dinhof & Walter Berger

  4. Laboratory for Cell Death Research and Therapy (CDRT), Department of Cellular and Molecular Medicine, KU Leuven University, Leuven, Belgium

    Abhishek D. Garg

  5. Dipartimento di Agraria, Sezione di Patologia vegetale ed Entomologia, Università degli Studi di Sassari, Viale Italia 39, 07100, Sassari, Italy

    Lucia Maddau

  6. Dipartimento di Scienze Chimiche, Universita’ di Napoli Federico II, Complesso Universitario Monte S. Angelo, Via Cintia 4, 80126, Napoli, Italy

    Marco Masi & Antonio Evidente

  7. Laboratoire d’Analyse Instrumentale et de Bioélectrochimie, Université Libre de Bruxelles, Boulevard du Triomphe, Accès 2, 1050, Ixelles, Belgium

    Bieke Dejaegher

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  1. Aude Ingels
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  2. Carina Dinhof
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  3. Abhishek D. Garg
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  4. Lucia Maddau
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  5. Marco Masi
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  6. Antonio Evidente
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  9. Véronique Mathieu
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Correspondence to Véronique Mathieu.

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Ingels, A., Dinhof, C., Garg, A.D. et al. Computed determination of the in vitro optimal chemocombinations of sphaeropsidin A with chemotherapeutic agents to combat melanomas. Cancer Chemother Pharmacol 79, 971–983 (2017). https://doi.org/10.1007/s00280-017-3293-x

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