5-Fluorocytosine increases the toxicity of Wnt-targeting replicating adenoviruses that express cytosine deaminase as a late gene.
Clinical studies with oncolytic adenoviruses have shown that existing viruses are safe but lack efficacy. To selectively increase the toxicity of oncolytic adenoviruses targeting colon tumours, we have inserted the yeast cytosine deaminase gene (yCD) after the fibre gene in the major late transcript. yCD was expressed using either an internal ribosome entry site (IRES) or by alternative splicing of a new exon analogous to the Ad41 long fibre exon. The IRES-CD virus gave higher yCD expression on Western blots. Both approaches result in yCD expression restricted to the period after viral DNA replication. Viral burst size was reduced by less than approximately 10-fold by 5-fluorocytosine (5-FC), showing that expression of yCD as a late gene is compatible with virus replication. Cytopathic effect assays in colon cancer cell lines showed that both yCD viruses have approximately 10-fold increased toxicity in the presence of the prodrug 5-FC, which is converted to 5-fluorouracil (5-FU) by yCD. Toxicity was higher following addition of 5-FC immediately after infection. The largest gain in toxicity was seen in HT29 colon cancer cells, which are the least permissive colon cancer cells for the parental virus, indicating that the new 5-FC/yCD viruses may have broader applications for colon cancer therapy than their predecessors.
Fuerer
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Oncogene Group, NCCR Molecular Oncology, Swiss Institute for Experimental Cancer Research (ISREC), Epalinges, Switzerland.
Iggo
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D000256:Adenoviridae / Q000235:genetics; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000378:metabolism / Q000628:therapy*; D043525:Cytosine Deaminase / Q000235:genetics; D005437:Flucytosine / Q000627:therapeutic use*; D015870:Gene Expression; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage; D019073:HT29 Cells; D006801:Humans; D011355:Prodrugs / Q000627:therapeutic use*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D014158:Transcription, Genetic; D014779:Virus Replication; D051153:Wnt Proteins; D029961:Zebrafish Proteins
D011355:Prodrugs; D011518:Proto-Oncogene Proteins; D051153:Wnt Proteins; D029961:Zebrafish Proteins; D005437:Flucytosine; D043525:Cytosine Deaminase