Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2019 Feb 15;12(1):18.
doi: 10.1186/s13048-019-0493-5.

Defining a murine ovarian cancer model for the evaluation of conditionally-replicative adenovirus (CRAd) virotherapy agents

Rebeca González-Pastor  1 , Ahmad Mohammad Ashshi  2 , Adel Galal El-Shemi  2   3 , Igor P Dmitriev  1 , Elena A Kashentseva  1 , Zhi Hong Lu  1 , S Peter Goedegebuure  4   5 , Osvaldo L Podhajcer  6 , David T Curiel  7
Affiliations

Defining a murine ovarian cancer model for the evaluation of conditionally-replicative adenovirus (CRAd) virotherapy agents

Rebeca González-Pastor et al. J Ovarian Res. .

Abstract

Background: Virotherapy represents a promising approach for ovarian cancer. In this regard, conditionally replicative adenovirus (CRAd) has been translated to the context of human clinical trials. Advanced design of CRAds has sought to exploit their capacity to induce anti-tumor immunization by configuring immunoregulatory molecule within the CRAd genome. Unfortunately, employed murine xenograft models do not allow full analysis of the immunologic activity linked to CRAd replication.

Results: We developed CRAds based on the Ad5/3-Delta24 design encoding cytokines. Whereas the encoded cytokines did not impact adversely CRAd-induced oncolysis in vitro, no gain in anti-tumor activity was noted in immune-incompetent murine models with human ovarian cancer xenografts. On this basis, we explored the potential utility of the murine syngeneic immunocompetent ID8 ovarian cancer model. Of note, the ID8 murine ovarian cancer cell lines exhibited CRAd-mediated cytolysis. The use of this model now enables the rational design of oncolytic agents to achieve anti-tumor immunotherapy.

Conclusions: Limits of widely employed murine xenograft models of ovarian cancer limit their utility for design and study of armed CRAd virotherapy agents. The ID8 model exhibited CRAd-induced oncolysis. This feature predicate its potential utility for the study of CRAd-based virotherapy agents.

Keywords: Adenovirus; Anti-tumor immunization; CRAd; ID8; Ovarian cancer; Virotherapy.

PubMed Disclaimer

Conflict of interest statement

Ethics approval and consent to participate

Not applicable.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no financial and non-financial competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Analysis of tumor growth inhibition following armed CRAd administration. Subcutaneous tumor xenografts were established on rear flank of female nude mice using SKOV3.ip1 cells. Tumor nodules were directly injected with 1010 vp (5 × 108 pfu/dose) of either CRAd-IL24, CRAd-ING4, CRAd-IL24/ING4 (5 × 109 vp of each armed CRAd), control CRAd or PBS alone on day 7 and injections were repeated weekly for 3 consecutive weeks. a Mean tumor volume of each group is shown through day 35 after tumor implantation. Each data point represents the cumulative mean of tumor volumes (mm3) in each group while vertical error bars depict standard deviations. b Kaplan-Meier curves of overall survival of animals treated with the indicated armed CRAd vectors, non-armed CRAd control, or PBS are shown. Data analysis provides no evidence that survival experience observed in mice treated with either CRAd-IL24, CRAd-ING4, or CRAd-IL24/ING4 (the indicated armed CRAd vectors) is significantly different from non-armed CRAd control (n.s.)
Fig. 2
Fig. 2
Analysis of cytopathic effects of replication competent Ad vectors. a Cytotoxic effects of wild type Ad5, Ad5-based Delta-24 and Delta-24-RGD CRAd vectors were compared in established ovarian cancer cell lines of human (SKOV3ip.1) and mouse (ID8 and ID8luc) origin using CellTox™ assayat the MOI of 100 vp/cell as compared to mock-infected cells (negative control) and completely lysed cells (positive assay control). Each data point represents the three replicate experiments ± SD (error bars are smaller than the symbols, p ≤ 0.05 *). b Monolayers of SKOV3ip.1, ID8, and ID8luc were infected with indicated Ad vectors and cell viability determined on day 6 post-infection by adding CellTiter 96 AQueous One Solution Reagent. The percentages of live cells in the monolayers exposed to replication-deficient E1-deleted Ad5∆E1 are shown as compared to wild type Ad5, Delta-24, and Delta-24-RGD-infected cells
Fig. 3
Fig. 3
Analysis of cytopathic effects of replication competent Ad vectors. a Cytotoxic effects of wild type Ad5 were compared in A549, GL261 and F3 using CellTox™ assay. Each data point represents the cumulative mean of triplicate measurements ± SD (p ≤ 0.05 *). b Monolayers of A549, GL261 and F3 were infected with indicated Ad vectors and cell viability measured after 5 days by adding CellTiter 96 AQueous One Solution Reagent. The percentages of live cells in the monolayers exposed to replication-deficient E1-deleted Ad5∆E1 are shown as compared to wild type Ad5-infected cells. c Replication was assed 48 h after infection at MOI 100vp/cell and 1000vp/cell using the Adeno-X-Rapid Titer kit. Final titers were determined at pfu/mL (p ≤ 0.05 *)
Fig. 4
Fig. 4
Analysis of tumor growth inhibition on a syngeneic orthotopic model following Ad administration. C57Bl/6 mice were injected i.p. with 5 × 106 F3mCherryLuc cells. After one week, mice were injected i.p. with 1 × 1010 vp of wild-type Ad or replication-incompetent Ad vectorsBLI images and corresponding signal quantification 6 days after Ad administration. Significant differences were found between PBS and AdWT groups (p = 0.0001 ****)
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Park JW, Kim M. Replicating viruses for gynecologic cancer therapy. Eur J Gynaecol Oncol. 2016;37(3):295–304. - PubMed
    1. Hartkopf AD, Fehm T, Wallwiener D, Lauer U. Oncolytic virotherapy of gynecologic malignancies. Gynecol Oncol. 2011;120(2):302–310. - PubMed
    1. Chan WM, Rahman MM, McFadden G. Oncolytic myxoma virus: the path to clinic. Vaccine. 2013;31(39):4252–4258. - PMC - PubMed
    1. Heiber JF, Xu XX, Barber GN. Potential of vesicular stomatitis virus as an oncolytic therapy for recurrent and drug-resistant ovarian cancer. Chin J Cancer. 2011;30(12):805–814. - PMC - PubMed
    1. Matthews KS, Alvarez RD, Curiel DT. Advancements in adenoviral based virotherapy for ovarian cancer. Adv Drug Deliv Rev. 2009;61(10):836–841. - PubMed

Substances