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HNF1β drives glutathione (GSH) synthesis underlying intrinsic carboplatin resistance of ovarian clear cell carcinoma (OCCC)

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Tumor Biology

Abstract

Chemoresistance to platinum-based antineoplastic agents is a consistent feature among ovarian carcinomas; however, whereas high-grade serous carcinoma (OSC) acquires resistance during chemotherapy, ovarian clear cell carcinoma (OCCC) is intrinsically resistant. The main objective of this study was to explore, in vitro and in vivo, if hepatocyte nuclear factor 1β (HNF1β) and glutaminolysis contribute for the resistance of OCCC to carboplatin through the intrinsically increased GSH bioavailability. To disclose the role of HNF1β, experiments were also performed in an OSC cell line, which does not express HNF1β. Metabolic profiles, GSH quantification, HNF1β, and γ-glutamylcysteine ligase catalytic subunit (GCLC) and modifier subunit (GCLM) expression, cell cycle, and death were assessed in ES2 cell line (OCCC) and OVCAR3 cell line (OSC); HNF1β knockdown was performed in ES2 and murine model of subcutaneous and peritoneal OCCC tumors was established to test buthionine sulphoxamine (BSO), as a sensitizer to carboplatin. Glutaminolysis is activated in ES2 and OVCAR3, though ES2 exclusively synthesizes amino acids and GSH. ES2 cells are more resistant to carboplatin than OVCAR3 and the abrogation of GSH production by BSO sensitizes ES2 to carboplatin. HNF1β regulates the expression of GCLC, but not GCLM, and consequently GSH production in ES2. In vivo, BSO prior to carboplatin reduces dramatically subcutaneous tumor size and GSH levels, as well as peritoneal dissemination. Our study discloses HNF1β as the mediator of intrinsic OCCC chemoresistance and sheds a light to re-explore a cancer adjuvant therapeutic approach using BSO to overcome the lack of efficient therapy in OCCC.

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Acknowledgments

The authors would like to thank to Instituto Português de Oncologia de Lisboa, Francisco Gentil, EPE (IPOLFG, EPE) for partially supporting the project, as well as to TVI in the context of Familial and individual cancer risk project. The NMR spectrometers are part of the National NMR Facility supported by Fundação para a Ciência e a Tecnologia (RECI/BBB-BQB/0230/2012). Sofia Gouveia-Fernandes acknowledges the fellowship from Liga Portuguesa Contra o Cancro and Roche Portugal (LPCC-Roche 2012). Luis G. Gonçalves acknowledges the fellowship from the project RECI/BBB-BQB/0230/2012 of Fundação para a Ciência e a Tecnologia. The authors would like to thank Duarte Barral (PhD; CEDOC-NOVA Medical School) for providing the access to the animal facility from Instituto Gulbenkian de Ciência, Oeiras.

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Authors and Affiliations

  1. Centro de Estudos de Doenças Crónicas (CEDOC), NOVA Medical School/Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Campo Mártires da Pátria 130, 1169-056, Lisbon, Portugal

    Filipa Lopes-Coelho, Sofia Gouveia-Fernandes, Carolina Nunes, Inês Faustino, Fernanda Silva, Ana Félix, Sofia A Pereira & Jacinta Serpa

  2. Unidade de Investigação em Patobiologia Molecular do Instituto Português de Oncologia de Lisboa Francisco Gentil (IPOLFG), Lisbon, Portugal

    Filipa Lopes-Coelho, Sofia Gouveia-Fernandes, Carolina Nunes, Fernanda Silva & Jacinta Serpa

  3. Instituto de Tecnologia Química e Biológica (ITQB), Oeiras, Portugal

    Luís G Gonçalves

  4. Pathology Department, IPOLFG, Lisbon, Portugal

    Ana Félix

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  1. Filipa Lopes-Coelho
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Correspondence to Jacinta Serpa.

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Supplementary Figure 1

BSO exposure reduces CYS-GLY and GSH levels in ES2 and OVCAR3. (JPEG 29 kb)

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Lopes-Coelho, F., Gouveia-Fernandes, S., Gonçalves, L.G. et al. HNF1β drives glutathione (GSH) synthesis underlying intrinsic carboplatin resistance of ovarian clear cell carcinoma (OCCC). Tumor Biol. 37, 4813–4829 (2016). https://doi.org/10.1007/s13277-015-4290-5

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