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.
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 x HRE) was cloned upstream of the sFlk-1 gene (10 x HRE/sFlk-1). A novel HSV amplicon expressing 10 x HRE/sFlk-1 was genetically engineered (HSV10 x HRE/sFlk-1).Human pancreatic adenocarcinoma cells (AsPC1) were transduced with HSV10 x 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 x HRE/sFlk-1.


RESULTS
Media from normoxic AsPC1 transduced with HSV10 x 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 x HRE/sFlk-1 demonstrated greater sFlk-1 expression in hypoxia versus normoxia. AsPC1 flank tumors treated with HSV10 x 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.
Reinblatt
Maura
M
Department of Surgery, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021, USA.
Pin
Richard H
RH
Bowers
William J
WJ
Federoff
Howard J
HJ
Fong
Yuman
Y
D000230:Adenocarcinoma / Q000473:pathology / Q000628:therapy*; D020533:Angiogenesis Inhibitors / Q000008:administration & dosage*; D015153:Blotting, Western; D015687:Cell Hypoxia; D004742:Enhancer Elements, Genetic; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051793:Hypoxia-Inducible Factor 1; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D010190:Pancreatic Neoplasms / Q000473:pathology / Q000628:therapy*; D018139:Simplexvirus / Q000235:genetics; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000008:administration & dosage*
D020533:Angiogenesis Inhibitors; D051793:Hypoxia-Inducible Factor 1; D040301:Vascular Endothelial Growth Factor Receptor-2