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Investigational New Drugs

, Volume 30, Issue 2, pp 568–581 | Cite as

Enhanced oncolysis mediated by Coxsackievirus A21 in combination with doxorubicin hydrochloride

  • Kathryn A. Skelding
  • Richard D. Barry
  • Darren R. ShafrenEmail author
PRECLINICAL STUDIES

Summary

Virotherapy is an emerging strategy for the treatment of cancer that utilizes both replication-competent and genetically modified viruses to selectively kill tumor cells. We have previously shown that Coxsackievirus A21 (CVA21), a common-cold producing enterovirus, is an effective oncolytic agent against human melanoma, prostate, and breast cancer xenografts in vivo. CVA21 specifically targets and lytically infects susceptible cells expressing the CVA21 cellular receptors, intercellular adhesion molecule-1 (ICAM-1) and decay-accelerating factor (DAF). Herein, the efficacy of CVA21 administered in combination with doxorubicin hydrochloride as a new therapeutic regimen for cancer was investigated. Flow cytometric analysis demonstrated that the human breast, colorectal, and pancreatic cancer cell lines examined expressed moderate levels of surface ICAM-1 and DAF, whilst a normal breast cell line expressed only minimal levels. When CVA21 was combined with doxorubicin hydrochloride, synergistically enhanced cell death was observed when CVA21 was administered both simultaneously or 24 h prior to doxorubicin hydrochloride exposure. Doxorubicin hydrochloride had no effect on CVA21 replication. Through the use of an orthotopic (MDA-MB-231-luc) xenograft SCID mouse model of human breast cancer we showed that a single intravenous injection of CVA21 in combination with an intraperitoneal injection of doxorubicin hydrochloride resulted in significantly greater tumor reduction compared to either agent alone. Overall, these findings highlight the exciting potential of CVA21, administered in combination with doxorubicin hydrochloride, as a new therapeutic regimen for cancer.

Keywords

Coxsackievirus A21 Oncolytic virus Breast cancer Colorectal cancer Pancreatic cancer Doxorubicin hydrochloride 

Abbreviations

ANOVA

One-way analysis of variance

CI

Combination index

CVA21

Coxsackievirus A21

DAF

Decay-accelerating factor

DMSO

Dimethyl sulfoxide

FBS

Fetal bovine serum

5-FU

5-fluorouracil

HSV-1

Herpes Simplex Virus-1

ICAM-1

Intercellular adhesion molecule-1

MOI

Multiplicity of infection

PBS

Phosphate buffered saline

SCID

Severe combined immunodeficient

SEM

Standard error of the mean

SPF

Specific-pathogen-free

TCID50

50% tissue culture infective dose

UV

Ultraviolet

Notes

Acknowledgements

We gratefully acknowledge those investigators mentioned in the text for the provision of monoclonal antibodies, cell lines and virus that enabled this study to be undertaken. We would like to thank Dr Gough Au, Dr Susanne Johansson, Dr Linda Berry, Dr Erin Haley, and Ms Catherine Delahunty for scientific advice and technical assistance throughout this project. Funding for this work was derived from grants received from the Biotechnology Innovation Fund and Hunter Medical Research Institute. Kathryn Skelding is a National Breast Cancer Foundation Scholar.

Declaration of competing interests

Darren Shafren and Richard Barry hold shares in Viralytics Ltd and are currently conducting research sponsored by this company.

Authors’ contributions

KS participated in the design of the study, carried out the in vitro and in vivo work, and drafted the manuscript. RB participated in the design of the study. DS participated in the design of the study, and was involved in the revision of the manuscript. All authors approved the final manuscript.

Supplementary material

10637_2010_9614_Fig9_ESM.gif (56 kb)
Supplementary Table 1

Combination index for MDA-MB-231 cells simultaneously treated with a combination of CVA21 and paclitaxel, tamoxifen, or 5-FU. Combination indices (CI) were calculated using CalcuSyn, as described in the Materials and methods. CI values less than 0.9 were determined to be synergistic, between 0.9 and 1.1 to be additive, and greater than 1.1 to be antagonistic. (GIF 56 kb)

10637_2010_9614_MOESM1_ESM.tif (9.4 mb)
High resolution image (TIFF 9597 kb)
10637_2010_9614_Fig10_ESM.gif (41 kb)
Supplementary Figure 1

Effect of doxorubicin hydrochloride, paclitaxel, 5-FU, and tamoxifen on CVA21 replication. MDA-MB-231 cell monolayers were inoculated with CVA21 (MOI = 10 TCID50/cell). Following the removal of unbound virus, the cell monolayers were incubated with (A) 2.5 or 0.32 μM doxorubicin hydrochloride, (B) 8 or 2 nM paclitaxel, (C) 10 nM or 10 μM 5-FU, or (D) 10 μM or 313 nM tamoxifen for 48 h. At selected time points (0, 2, 4, 6, 8, 10, 24, and 48 h), cell suspensions were freeze-thawed, clarified, and virus yield was determined by viral lytic assay on SK-Mel-28 cells. (GIF 40 kb)

10637_2010_9614_MOESM2_ESM.tif (2.1 mb)
High resolution image (TIFF 2140 kb)

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Kathryn A. Skelding
    • 1
  • Richard D. Barry
    • 1
  • Darren R. Shafren
    • 1
    • 2
    Email author
  1. 1.The Picornaviral Research Unit, Discipline of Immunology and Microbiology, Faculty of HealthThe University of NewcastleNewcastleAustralia
  2. 2.Viralytics LtdPymbleAustralia

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