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Ex vivo-expanded NK cells from blood and ascites of ovarian cancer patients are cytotoxic against autologous primary ovarian cancer cells

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Abstract

Ovarian cancer (OC) is the leading cause of gynecological cancer-related death in North America. Most ovarian cancer patients (OCPs) experience disease recurrence after first-line surgery and chemotherapy; thus, there is a need for novel second-line treatments to improve the prognosis of OC. Although peripheral blood-derived NK cells are known for their ability to spontaneously lyse tumour cells without prior sensitization, ascites-derived NK cells (ascites-NK cells) isolated from OCPs exhibit inhibitory phenotypes, impaired cytotoxicity and may play a pro-tumourigenic role in cancer progression. Therefore, it is of interest to improve the cytotoxic effector function of impaired OCP ascites-NK cells at the tumour environment. We investigated the efficacy of using an artificial APC-based ex vivo expansion technique to generate cytotoxic, expanded NK cells from previously impaired OCP ascites-NK cells, for use in an autologous model of NK cell immunotherapy. We are the first to obtain a log-scale expansion of OCP ascites-NK cells that upregulate the surface expression of activating receptors NKG2D, NKp30, NKp44, produce robust amounts of anti-tumour cytokines in the presence of OC cells and mediate direct tumour cytotoxicity against ascites-derived, primary OC cells obtained from autologous patients. Our findings demonstrate that it is possible to generate cytotoxic OCP ascites-NK cells from previously impaired OCP ascites-NK cells, which presents a promising immunotherapeutic target for the second-line treatment of OC. Future work should focus on evaluating the in vivo efficacy of autologous NK cell immunotherapy through the intraperitoneal delivery of NK cell expansion factors to a preclinical xenograft mouse model of human OC.

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Abbreviations

aAPC:

Artificial antigen-presenting cell

CFSE:

Carboxyfluorescein succinimidyl ester

E:T:

Effector: target

HD:

Healthy donor

IFN-γ:

Interferon-γ

IL:

Interleukin

NK cells:

Natural killer cells

OC:

Ovarian cancer

OCP:

Ovarian cancer patient

OCP ascites-NK cells:

NK cells derived from ovarian cancer patient ascites

PB:

Peripheral blood

PB-NK cells:

NK cells derived from peripheral blood

TNF-α:

Tumor necrosis factor-α

VEGF:

Vascular endothelial growth factor

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Acknowledgements

We thank Dr. Karen Mossman for providing us with the OVCAR-8 cell line. We also thank Jann Ang and Natasha Kazhdan for aiding us in sample collection.

Funding

Funding for this research was supported by grants from the Juravinski Hospital and Cancer Centre Foundation, the Canadian Breast Cancer Foundation and the Canadian Institutes of Health Research Tier 1 Canada Research Chair awarded to Dr. Ali A. Ashkar.

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Authors and Affiliations

  1. Department of Pathology and Molecular Medicine, McMaster Immunology Research Centre, McMaster University, Rm 4015 Michael DeGroote Centre for Learning and Discovery, 1280 Main Street West, Hamilton, ON, L8S 4K1, Canada

    Tina Nham, Sophie M. Poznanski, Isabella Y. Fan, Mira M. Shenouda, Marianne V. Chew, Amanda J. Lee, Fatemeh Vahedi, Yalda Karimi & Ali A. Ashkar

  2. Juravinski Cancer Centre, McMaster University, Hamilton, ON, Canada

    Martin Butcher

  3. The Research Institute at Nationwide Children’s Hospital, Ohio State University, Columbus, OH, USA

    Dean A. Lee

  4. Division of Medical Oncology, Department of Oncology, Juravinski Cancer Centre, Hamilton, ON, Canada

    Hal Hirte

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  1. Tina Nham
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  2. Sophie M. Poznanski
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Correspondence to Ali A. Ashkar.

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The authors declare that they have no conflict of interest.

Cell line authentication

The OVCAR-8 cell line was verified using the American Type Culture Collection® (ATCC®) Cell Line Authentication Service for which we received a Short Tandem Repeat Profile Report on September 5, 2017. The submitted OVCAR-8 sample is an exact match to the short tandem repeat profile for the cell line OVCAR-8 listed on the Expasy website: http://web.expasy.org/cellosaurus/CVCL_1629.

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Nham, T., Poznanski, S.M., Fan, I.Y. et al. Ex vivo-expanded NK cells from blood and ascites of ovarian cancer patients are cytotoxic against autologous primary ovarian cancer cells. Cancer Immunol Immunother 67, 575–587 (2018). https://doi.org/10.1007/s00262-017-2112-x

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