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Context-dependent activation of SIRT3 is necessary for anchorage-independent survival and metastasis of ovarian cancer cells

Oncogene volume 39, pages 1619–1633 (2020)Cite this article

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Abstract

Tumor cells must alter their antioxidant capacity for maximal metastatic potential. Yet the antioxidant adaptations required for ovarian cancer transcoelomic metastasis, which is the passive dissemination of cells in the peritoneal cavity, remain largely unexplored. Somewhat contradicting the need for oxidant scavenging are previous observations that expression of SIRT3, a nutrient stress sensor and regulator of mitochondrial antioxidant defenses, is often suppressed in many primary tumors. We have discovered that this mitochondrial deacetylase is specifically upregulated in a context-dependent manner in cancer cells. SIRT3 activity and expression transiently increased following ovarian cancer cell detachment and in tumor cells derived from malignant ascites of high-grade serous adenocarcinoma patients. Mechanistically, SIRT3 prevents mitochondrial superoxide surges in detached cells by regulating the manganese superoxide dismutase (SOD2). This mitochondrial stress response is under dual regulation by SIRT3. SIRT3 rapidly increases SOD2 activity as an early adaptation to cellular detachment, which is followed by SIRT3-dependent increases in SOD2 mRNA during sustained anchorage-independence. In addition, SIRT3 inhibits glycolytic capacity in anchorage-independent cells thereby contributing to metabolic changes in response to detachment. While manipulation of SIRT3 expression has few deleterious effects on cancer cells in attached conditions, SIRT3 upregulation and SIRT3-mediated oxidant scavenging are required for anoikis resistance in vitro following matrix detachment, and both SIRT3 and SOD2 are necessary for colonization of the peritoneal cavity in vivo. Our results highlight the novel context-specific, pro-metastatic role of SIRT3 in ovarian cancer.

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Acknowledgements

We thank Usawadee Dier, Dr LP Madhubhani Hemachandra, Dr Sarah Engelberth, Larissa Suparmanto, Sadie Dierschke, and Gina Deiter for technical assistance. We thank Dr Arati Sharma and Dr David Claxton (Penn State) for providing PDX animals. We are grateful for Dr Vonn Walter’s advice on TCGA data analysis. MnTnBuOE-2-PyP5+ was kindly provided by Dr Ines Batinic Haberle (Duke University). OVCA433 and OVCA420 cells were a generous gift from Dr Susan Murphy (Duke University). This work was supported by NIH grants R00CA143229 (NH), R01CA230628 (NH & KM), S100D018124 (TA), by the Rivkin Center for Ovarian Cancer (NH), an equipment grant from Seahorse/Agilent (NH), and the Penn State Cancer Institute Developmental Fund Award (NH). Collection of ascites was partially supported by DoD Pilot award W81XWH-16-1-0117 (NH).

Funding

This work was supported by NIH grants R00CA143229 (NH), R01CA230628 (NH and KM), S100D018124 (TA), by the Rivkin Center for Ovarian Cancer (NH), an equipment grant from Agilent (NH), and the Penn State Cancer Institute Developmental Fund Award (NH). Collection of ascites was partially supported by DoD Pilot award W81XWH-16-1-0117 (NH).

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Author notes

  1. These authors contributed equally: Yeon Soo Kim, Piyushi Gupta Vallur

Authors and Affiliations

  1. Department of Pharmacology, College of Medicine, Pennsylvania State University, Hershey, PA, USA

    Yeon Soo Kim, Piyushi Gupta Vallur, Victoria M. Jones, Beth L. Worley, Sara Shimko, Dong-Hui Shin, LaTaijah C. Crawford & Nadine Hempel

  2. Department of Cellular and Molecular Physiology, College of Medicine, Pennsylvania State University, Hershey, PA, USA

    Chi-Wei Chen & Katherine M. Aird

  3. Department of Neural and Behavioral Sciences, College of Medicine, Pennsylvania State University, Hershey, PA, USA

    Thomas Abraham

  4. The Mary & John Knight Translational Ovarian Cancer Research Unit, Departments of Obstetrics & Gynecology Oncology and Anatomy & Cell Biology, Western University, London, ON, Canada

    Trevor G. Shepherd

  5. Department of Pathology, College of Medicine, Pennsylvania State University, Hershey, PA, USA

    Joshua I. Warrick

  6. Department of Pharmacology, College of Medicine, University of Arizona, Tucson, AZ, USA

    Nam Y. Lee

  7. Department of Obstetrics and Gynecology, and Microbiology and Immunology, College of Medicine, Pennsylvania State University, Hershey, PA, USA

    Rebecca Phaeton

  8. Department of Chemistry and Biochemistry, University of South Carolina, Columbia, SC, USA

    Karthikeyan Mythreye

  9. Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA

    Karthikeyan Mythreye

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  1. Yeon Soo Kim
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Contributions

YSK and PGV contributed to study design, manuscript writing, and the majority of experimental execution and data analysis. VMJ and DHS assisted with cell culture studies. LCC carried out Seahorse experiments. BLW and SS contributed to in vivo studies and data analysis. CWC and KMA performed media glucose and lactate quantification. TA assisted in multiphoton experiments and performed data analysis. TGS carried out microarray expression and data analysis. JIW performed analysis of tumor sections. NYL assisted in data interpretation and manuscript editing. RP provided patient ascites and tumor cells and assisted in study design. KM contributed to conceptual design, data interpretation and writing of the manuscript. NH conceived and supervised the study, designed experiments and wrote the manuscript.

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Correspondence to Karthikeyan Mythreye or Nadine Hempel.

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Kim, Y.S., Gupta Vallur, P., Jones, V.M. et al. Context-dependent activation of SIRT3 is necessary for anchorage-independent survival and metastasis of ovarian cancer cells. Oncogene 39, 1619–1633 (2020). https://doi.org/10.1038/s41388-019-1097-7

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