Abstract

The efficacy of oncolytic herpes simplex virus (oHSV) is limited by rapid viral clearance by innate immune effector cells and poor intratumoral viral spread. We combine two approaches to overcome these barriers: inhibition of natural killer (NK) cells and enhancement of intratumoral viral spread. We engineered an oHSV to express CDH1, encoding E-cadherin, an adherent molecule and a ligand for KLRG1, an inhibitory receptor expressed on NK cells. In vitro, infection with this engineered virus, named OV-CDH1, induced high surface E-cadherin expression on infected glioblastoma (GBM) cells, which typically lack endogenous E-cadherin. Ectopically expressed E-cadherin enhanced the spread of OV-CDH1 by facilitating cell-to-cell infection and viral entry and reduced viral clearance by selectively protecting OV-CDH1-infected cells from KLRG1+ NK cell killing. In vivo, OV-CDH1 treatment substantially prolonged the survival in GBM-bearing mouse models, primarily because of improved viral spread rather than inhibition of NK cell activity. Thus, virus-induced overexpression of E-cadherin may be a generalizable strategy for improving cancer virotherapy.

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Acknowledgements

The authors thank A. Yilmaz, B. McNeil and V. Sellers for critical reading. This work was supported by grants from the NIH (NS106170, AI129582, J.Y.; CA185301, CA068458, CA210087, CA163205-5805, M.A.C.; CA163205-5806, E.A.C.), the Leukemia & Lymphoma Society (6503-17, 1364-19, J.Y.), the American Cancer Society (RSG-14-243-01-LIB, J.Y.) and the Gabrielle's Angel Cancer Research Foundation (87, J.Y.).

Author information

Author notes

    • Bo Xu
    •  & Rui Ma

    These authors contributed equally to this work.

Affiliations

  1. Division of Hematology, Department of Internal Medicine, College of Medicine, The Ohio State University, Columbus, Ohio, USA.

    • Bo Xu
    •  & Jianhua Yu
  2. The Ohio State University Comprehensive Cancer Center, Columbus, Ohio, USA.

    • Bo Xu
    • , Rui Ma
    • , Luke Russell
    • , Jianfeng Han
    • , Hanwei Cui
    • , Ping Yi
    • , Hongsheng Dai
    •  & Jianhua Yu
  3. Department of Neurosurgery, McGovern Medical School, University of Texas Health Science Center at Houston, Houston, Texas, USA.

    • Ji Young Yoo
    •  & Balveen Kaur
  4. Third Affiliated Hospital, Army Medical University, Chongqing, China.

    • Hanwei Cui
    •  & Ping Yi
  5. Division of Biostatistics, Department of Information Sciences, City of Hope National Medical Center, Duarte, California, USA.

    • Jianying Zhang
  6. Department of Neurosurgery, Brigham and Women's Hospital and Harvey Cushing Neuro-oncology Laboratories, Harvard Medical School, Boston, Massachusetts, USA.

    • Hiroshi Nakashima
    •  & E Antonio Chiocca
  7. Hematologic Malignancies and Stem Cell Transplantation Institute, City of Hope National Medical Center, Duarte, California, USA.

    • Michael A Caligiuri
    •  & Jianhua Yu
  8. Department of Hematology & Hematopoietic Cell Transplantation, City of Hope National Medical Center and Beckman Research Institute, Duarte, California, USA.

    • Michael A Caligiuri
    •  & Jianhua Yu

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Contributions

B.X. performed experiments, designed research and wrote the manuscript; R.M., L.R., J.Y.Y. and J.H., H.C., P.Y. performed experiments; H.D. designed research; J.Z. analyzed the data; H.N. provided materials; E.A.C. and B.K. designed research and reviewed the manuscript. M.A.C. designed research, reviewed and edited the manuscript and acquired funding. J.Y. designed research, wrote the manuscript, acquired funding and supervised the study.

Competing interests

A patent application on OV-CDH1 virus related to this work has been submitted by The Ohio State University on behalf of the inventors, J.Y., M.A.C. and B.X.

Corresponding author

Correspondence to Jianhua Yu.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–19, Supplementary Table 1

  2. 2.

    Life Sciences Reporting Summary

  3. 3.

    Supplementary Notes

    Supplementary Notes 1–4

Videos

  1. 1.

    Supplementary Video 1

    The plaque-forming process of OV-Q1-infected U251 cells. U251 cells were infected with OV-Q1 at a MOI of 0.005. At 2 hpi, infection media were replaced with fresh media. The video was recorded from 24 to 72 hpi using Zeiss fluorescence microscope (AXIO observer Z1). The time interval is 5 min. Green fluorescence (GFP) indicates virus-infected cells. This experiment was repeated 3 times with similar results.

  2. 2.

    Supplementary Video 2

    The plaque-forming process of OV-CDH1-infected U251 cells. U251 cells were infected with OV-CDH1 at a MOI of 0.005. At 2 hpi, infection media were replaced with fresh media. The video was recorded from 24 to 72 hpi using Zeiss fluorescence microscope (AXIO observer Z1). The time interval is 5 min. Green fluorescence (GFP) indicates virus-infected cells. This experiment was repeated 3 times with similar results.

  3. 3.

    Supplementary Video 3

    The plaque-forming process of OV-Q1-infected U251 cells after staining with CellTracker. U251 cells were infected with OV-Q1 at a MOI of 0.005. At 2 hpi, infection media were replaced with fresh media. At 24 hpi, cells were stained with Celltracker Deep Red. The video was recorded from 24 to 72 hpi using Zeiss fluorescence microscope (AXIO observer Z1). The time interval is 5 min. Green fluorescence (GFP) indicates virus-infected cells; red fluorescence indicates the cytoplasmic content of all the cells stained with CellTracker. This experiment was repeated 3 times with similar results.

  4. 4.

    Supplementary Video 4

    The plaque-forming process of OV-CDH1-infected U251 cells after staining with CellTracker. U251 cells were infected with OV-CDH1 at a MOI of 0.005. At 2 hpi, infection media were replaced with fresh media. At 24 hpi, cells were stained with Celltracker Deep Red. The video was recorded from 24 to 72 hpi using Zeiss fluorescence microscope (AXIO observer Z1). The time interval is 5 min. Green fluorescence (GFP) indicates virus-infected cells; red fluorescence indicates the cytoplasmic content of all the cells stained with CellTracker. This experiment was repeated 3 times with similar results.

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DOI

https://doi.org/10.1038/nbt.4302