Carcinoembryonic antigen directed herpes viral oncolysis improves selectivity and activity in colorectal cancer.
BACKGROUND
G207 is an oncolytic herpes virus whose replicative cycle requires cellular ribonucleotide reductase (RR) for viral DNA synthesis. We attempt to enhance viral cytotoxicity in carcinoembryonic antigen (CEA)-producing colorectal cancer (CRC) cells through CEA-driven RR production.


METHODS
CEA enzyme-linked immunosorbent assay was performed on LS174T and HCT-8 human CRC cells. The CEA enhancer-promoter (CEA E-P) was functionally assessed by luciferase assay. CEA E-P was cloned upstream of UL39, the gene encoding the large subunit of RR. Cells were transfected with CEA E-P/UL39 and infected with G207 for cytotoxicity assays. LS174T, with or without CEA E-P/UL39, were implanted into athymic mouse flanks (n = 28) and treated with G207.


RESULTS
CEA levels were 7-fold higher in LS174T versus HCT-8 ( P <.00001). CEA E-P increased luciferase expression 7.5-fold in LS174T ( P <.01), with no increase in HCT-8. G207 cytotoxicity of'CEA E-P/UL39-transfected LS174T cells increased 69% by day 10 versus nontransfected cells ( P <.001), with no significant increase in HCT-8. Combining CEA E-P/UL39 with G207 in LS174T flank tumors resulted in a 65% decrease in tumor volume versus G207, phosphate-buffered saline, or'CEA E-P/UL39 alone ( P <.0001).


CONCLUSIONS
CEA-driven RR production by CEA-secreting CRC cells significantly improves oncolytic viral cytotoxicity and specificity in vitro, and reduces tumor burden in vivo.
Reinblatt
Maura
M
Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, NY, USA.
Pin
Richard H
RH
Fong
Yuman
Y
D002272:Carcinoembryonic Antigen / Q000235:genetics*; D016923:Cell Death; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000628:therapy*; D015972:Gene Expression Regulation, Neoplastic; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D012264:Ribonucleotide Reductases / Q000502:physiology; D018139:Simplexvirus / Q000235:genetics*; D014779:Virus Replication / Q000502:physiology; D014780:Viruses / Q000235:genetics
D002272:Carcinoembryonic Antigen; D012264:Ribonucleotide Reductases