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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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.).
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.
Supplementary Figures 1–19, Supplementary Table 1 (PDF 7693 kb)
Supplementary Notes 1–4 (PDF 150 kb)
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. (AVI 30223 kb)
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. (AVI 44622 kb)
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. (AVI 26612 kb)
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. (AVI 40372 kb)
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Xu, B., Ma, R., Russell, L. et al. An oncolytic herpesvirus expressing E-cadherin improves survival in mouse models of glioblastoma. Nat Biotechnol 37, 45–54 (2019). https://doi.org/10.1038/nbt.4302
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