Optimizing oncolytic adenoviruses for treatment of cholangiocarcinoma

Chen, Yi-Hsuan (2014). Optimizing oncolytic adenoviruses for treatment of cholangiocarcinoma. University of Birmingham. M.Res.

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Abstract

Oncolytic adenoviruses offer a promising new treatment for cancers, especially those which respond poorly to current therapies such as cholangiocarcinoma. However, for clinical use, high selectivity to cancers is required. Thus, I constructed two EGFP-reporter gene expressing viruses, hTERTp-E1AWT-EGFP and E2Fp-E1AΔ24-EGFP to allow the comparison with an existing WTp-E1AΔ24-EGFP virus. These viruses were compared for their ability to infect and replicate in two cholangiocarcinoma cell lines, CCLP1 and CCSW1 cells.
Flow cytometry was used to monitor EGFP expression by the replicating viruses. In these experiments, the virus WTp-E1AΔ24 (virus B) reproducibly showed the greatest EGFP expression at equivalent multiplicity of infection. This replication efficacy was also confirmed in qPCR experiments measuring viral genome copy number. In cell viability assays, the hTERTp-E1AWT virus (virus A) was less potent than either E2Fp-E1AΔ24 (virus C) or WTp-E1AΔ24 (virus B), which had similar oncolytic potency.
I also compared the ability of replication-defective adenoviruses expressing EGFP and a range of alternative fibre proteins to infect the cholangiocarcinoma cell lines. Fibre proteins incorporating the knob domain from adenovirus type 3, or including the A20-RGD (arginine, glycine and aspartate)-containing peptide from foot and mouth disease virus, showed significantly improved infectivity. These data will assist in the development of improved oncolytic viruses for treatment of cholangiocarcinoma.

Type of Work:

Thesis (Masters by Research > M.Res.)

Award Type:

Masters by Research > M.Res.

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