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Herpes Simplex Virus Amplicon Delivery of a Hypoxia-Inducible Soluble Vascular Endothelial Growth Factor Receptor (sFlk-1) Inhibits Angiogenesis and Tumor Growth in Pancreatic Adenocarcinoma

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Abstract

Background

Tumor hypoxia induces vascular endothelial growth factor (VEGF) expression, which stimulates angiogenesis and tumor proliferation. The VEGF signaling pathway is inhibited by soluble VEGF receptors (soluble fetal liver kinase 1; sFlk-1), which bind VEGF and block its interaction with endothelial cells. Herpes simplex virus (HSV) amplicons are replication-incompetent viruses used for gene delivery. We attempted to attenuate angiogenesis and inhibit pancreatic tumor growth through HSV amplicon–mediated expression of sFlk-1 under hypoxic control.

Methods

A multimerized hypoxia-responsive enhancer (10 × HRE) was cloned upstream of the sFlk-1 gene (10 × HRE/sFlk-1). A novel HSV amplicon expressing 10 × HRE/sFlk-1 was genetically engineered (HSV10 × HRE/sFlk-1).Human pancreatic adenocarcinoma cells (AsPC1) were transduced with HSV10 × HRE/sFlk-1 and incubated in normoxia (21% oxygen) or hypoxia (1% oxygen). Capillary inhibition was evaluated by human umbilical vein endothelial cell assay. Western blot assessed sFlk-1 expression. AsPC1 flank tumor xenografts (n = 24) were transduced with HSV10 × HRE/sFlk-1.

Results

Media from normoxic AsPC1 transduced with HSV10 × HRE/sFlk-1 yielded a 36% reduction in capillary formation versus controls (P < .05), whereas hypoxic AsPC1 yielded a 76% reduction (P < .005). Western blot of AsPC1 transduced with HSV10 × HRE/sFlk-1 demonstrated greater sFlk-1 expression in hypoxia versus normoxia. AsPC1 flank tumors treated with HSV10 × HRE/sFlk-1 exhibited a 59% reduction in volume versus controls (P < .000001).

Conclusions

HSV amplicon delivery of a hypoxia-inducible soluble VEGF receptor significantly reduces new vessel formation and tumor growth. Tumor hypoxia can thus be used to direct antiangiogenic therapy to pancreatic adenocarcinoma.

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Acknowledgments

The authors thank Guo-jie Ye, PhD, for his advice with cloning procedures and knowledge of HSV vectors. They also thank Sam Yoon, MD, for his help with the HUVEC assay and Wade Narrows for preparation of helper virus-free HSV amplicon stocks. Supported by grants RO1 CA 76416 and RO1 CA/DK80982 (Y.F.) from the National Institutes of Health, grant MBC-99366 (Y.F.) from the American Cancer Society, grant BC024118 from the US Army, and the Rochester Nathan Shock Center.

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Authors and Affiliations

  1. Department of Surgery, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, New York, 10021

    Maura Reinblatt MD, Richard H. Pin MD & Yuman Fong MD

  2. Center for Aging and Developmental Biology, University of Rochester School of Medicine and Dentistry, Rochester, New York, 14642

    William J. Bowers PhD & Howard J. Federoff MD, PhD

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  1. Maura Reinblatt MD
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  2. Richard H. Pin MD
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  3. William J. Bowers PhD
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  5. Yuman Fong MD
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Correspondence to Yuman Fong MD.

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Reinblatt, M., Pin, R.H., Bowers, W.J. et al. Herpes Simplex Virus Amplicon Delivery of a Hypoxia-Inducible Soluble Vascular Endothelial Growth Factor Receptor (sFlk-1) Inhibits Angiogenesis and Tumor Growth in Pancreatic Adenocarcinoma. Ann Surg Oncol 12, 1025–1036 (2005). https://doi.org/10.1245/ASO.2005.03.081

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  • DOI: https://doi.org/10.1245/ASO.2005.03.081

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