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Phytoglycogen-dsRNA nanoparticles demonstrate differential cytotoxicity and immunostimulatory potential in two ovarian cancer cell lines

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Abstract

Ovarian cancer is a leading cause of cancer mortality in women, and novel treatments with improved efficacy are needed to fight ovarian cancer. Double-stranded (ds) RNA, including the synthetic polyinosinic cytidylic acid (poly (I:C), has shown promise as a cancer therapeutic. Phytoglycogen derived from sweet corn, nanodendrix (NDX) is a carrier for dsRNA. The responsiveness to NDX-delivered poly (I:C), NDX-poly (I:C), was tested in two ovarian cancer cell lines, SKOV-3 and OVCAR-3, previously identified as dsRNA-resistant and dsRNA-sensitive, respectively. NDX bound poly (I:C) at a w/w ratio of 2:1 NDX:poly (I:C) and poly (I:C)-NDX was tested for biological activity through uptake and two therapeutic modes of action, cytotoxicity, and immune stimulation. Immunocytochemistry demonstrated both cells bound to poly (I:C)-NDX. In OVCAR-3, poly (I:C)-NDX caused significant cell death, even at concentrations as low as 62.5 ng/mL; no cell death was observed with poly (I:C) alone at concentrations up to 5 μg/mL in SKOV-3 and 0.5 μg/mL in OVCAR-3. In both cell lines poly (I:C)-NDX stimulated the production of CXCL10 protein and transcripts, an innate immune chemokine, and at significantly higher levels than poly (I:C) alone. Interestingly, in response to poly (I:C)-NDX SKOV-3 produced a more robust immune response and higher levels of capase-3/-7 activation compared to OVCAR-3, despite showing no significant cell death. Poly (I:C)-NDX represents a robust and multifunctional therapy, potentiating poly (I:C) and sensitizing resistant cells. Additionally, the SKOV-3 and OVCAR-3 combination represents a powerful comparative model to help unravel dsRNA-mediated immune responses in ovarian cancer cells.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors would like to thank Jondavid de Jong for his mentorship and guidance.

Funding

This research was supported by the Wilfrid Laurier University (SP and SDO) and an NSERSC CRD (SDO).

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Authors and Affiliations

  1. Department of Health Sciences, Wilfrid Laurier University, 75 University Avenue West, Waterloo, ON, Canada

    A. Lewis, N. L. Aldor, N. A. Jadaa, S. J. DeWitte-Orr & S. J. Poynter

  2. Department of Biology, Wilfrid Laurier University, 75 University Avenue West, Waterloo, ON, Canada

    A. Tran & S. J. DeWitte-Orr

  3. Mirexus Biotechnologies Inc, 574 Hanlon Creek Boulevard, Guelph, ON, Canada

    T. Feng & E. Moore

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Contributions

Aaron Lewis completed experiments for Figs. 1, 3, and 4 and assisted in manuscript preparation. Andy Tran contributed to experiments for Fig. 2. Natalie Aldor contributed to experiments for Figs. 4 and 5 and assisted in manuscript preparation. Nicholas Jadaa contributed to experiments for Fig. 5. Tina Feng and Emily Moore synthesized the nanoparticle starting material. Stephanie DeWitte-Orr and Sarah Poynter developed the concept, designed experiments, and assisted in manuscript preparation.

Corresponding author

Correspondence to S. J. Poynter.

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Ethics approval was obtained from Wilfrid Laurier University for use of human cell lines (REB 6993)

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Conflict of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. EM and TF were paid employees of Mirexus Biotechnologies Inc. at the time of nanoparticle synthesis but were not involved in any tests using the nanoparticle in vitro. SP is a previous employee of Mirexus Biotechnologies Inc. but no longer has any financial connection to the company. SDO is an unpaid member of the Scientific Advisory Board for Mirexus Biotechnologies Inc. Authors AL, AT, NA, and NJ have no affiliation with Mirexus Biotechnologies Inc. As such all authors declare no competing financial or non-financial interests.

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Lewis, A., Tran, A., Aldor, N.L. et al. Phytoglycogen-dsRNA nanoparticles demonstrate differential cytotoxicity and immunostimulatory potential in two ovarian cancer cell lines. J Nanopart Res 25, 104 (2023). https://doi.org/10.1007/s11051-023-05758-7

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