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Adenoviral-mediated imaging of gene transfer using a somatostatin receptor-cytosine deaminase fusion protein

Cancer Gene Therapy volume 22, pages 215–221 (2015)Cite this article

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Abstract

Suicide gene therapy is a process by which cells are administered a gene that encodes a protein capable of converting a nontoxic prodrug into an active toxin. Cytosine deaminase (CD) has been widely investigated as a means of suicide gene therapy owing to the enzyme’s ability to convert the prodrug 5-fluorocytosine (5-FC) into the toxic compound 5-fluorouracil (5-FU). However, the extent of gene transfer is a limiting factor in predicting therapeutic outcome. The ability to monitor gene transfer, non-invasively, would strengthen the efficiency of therapy. In this regard, we have constructed and evaluated a replication-deficient adenovirus (Ad) containing the human somatostatin receptor subtype 2 (SSTR2) fused with a C-terminal yeast CD gene for the non-invasive monitoring of gene transfer and therapy. The resulting Ad (AdSSTR2-yCD) was evaluated in vitro in breast cancer cells to determine the function of the fusion protein. These studies demonstrated that both the SSTR2 and yCD were functional in binding assays, conversion assays and cytotoxicity assays. In vivo studies similarly demonstrated the functionality using conversion assays, biodistribution studies and small animal positron-emission tomography (PET) imaging studies. In conclusion, the fusion protein has been validated as useful for the non-invasive imaging of yCD expression and will be evaluated in the future for monitoring yCD-based therapy.

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Acknowledgements

This work was supported by the Department of Radiation Oncology, Washington University School of Medicine and NIH grant R01 EB004533. We gratefully acknowledge Margaret Morris, Nicole Fettig, Lori Strong and Amanda Roth for performing the small animal imaging studies. Dr. Jeffrey Craft is thanked for his assistance in analysis of the imaging data.

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Author notes

  1. T J Wadas

    Present address: 3Current Address: Department of Cancer Biology, Wake Forest University, Winston-Salem, NC, USA,

Authors and Affiliations

  1. Department of Radiation Oncology, Washington University School of Medicine, St Louis, MO, USA

    K A Lears, J J Parry, R Andrews, K Nguyen & B E Rogers

  2. Mallinckrodt Institute of Radiology, Washington University School of Medicine, St Louis, MO, USA

    T J Wadas & B E Rogers

Authors
  1. K A Lears
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Correspondence to B E Rogers.

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Lears, K., Parry, J., Andrews, R. et al. Adenoviral-mediated imaging of gene transfer using a somatostatin receptor-cytosine deaminase fusion protein. Cancer Gene Ther 22, 215–221 (2015). https://doi.org/10.1038/cgt.2015.14

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