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Detection and measurement of in vitro gene transfer by gamma camera imaging

Gene Therapy volume 8, pages 291–299 (2001)Cite this article

Abstract

The purpose of this work was to develop a high capacity method to image gene transfer to cancer cells growing as monolayers in cell culture plates. A sensitive and high capacity nuclear-imaging method for detection of gene transfer in vitro will allow rapid validation of vectors in different cell lines under various conditions. Human cancer cell lines (A-427 non-small cell lung, SKOV3.ip1 ovarian, MDA-MB-468 breast, and BxPC-3 pancreatic) were infected with a replication-incompetent adenoviral vector encoding the human type 2 somatostatin receptor (Ad-hSSTr2). Expression of the hSSTr2 reporter protein in cells was detected by imaging an internalized 99mTc-labeled, hSSTr2 binding peptide (P2045, Diatide, Inc.). Imaging provided an accurate measure of internally bound 99mTc as evidenced by equivalence of results for imaging region of interest (ROI) analyses and gamma counter measurements. Internally bound 99mTc-P2045 was linearly correlated (R2 = 0.98) with the percentage of hSSTr2-positive cells following gene transfer. Excess P2045 blocked binding and internalization of the 99mTc-P2045, indicating the specificity of the technique. Up to four 96-well plates could be imaged simultaneously, thereby demonstrating the high capacity of the system. This novel in vitro approach provides a new method to test enhanced gene transfer as new vectors are developed.

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Acknowledgements

This work was supported in part by NCI grant CA80104, NIH grant CA73636 and DOE grant DE-FG02-93ER61654.The authors greatly appreciate the generous supply of the P2045 from Diatide, Inc. The authors thank Gabriela Anchondo, Gloria Robinson, Debbie Della Manna, and Richard Kirkman for excellent technical assistance.

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Authors and Affiliations

  1. Department of Radiology, University of Alabama at Birmingham, Birmingham, AL, USA

    KR Zinn, TR Chaudhuri & JM Mountz

  2. Department of Radiation Oncology, University of Alabama at Birmingham, Birmingham, AL, USA

    DJ Buchsbaum & BE Rogers

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  1. KR Zinn
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Zinn, K., Chaudhuri, T., Buchsbaum, D. et al. Detection and measurement of in vitro gene transfer by gamma camera imaging. Gene Ther 8, 291–299 (2001). https://doi.org/10.1038/sj.gt.3301391

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