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

Gene Therapy
K R ZinnB E Rogers

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 ...Continue Reading

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Citations

Oct 18, 2001·Cancer Biotherapy & Radiopharmaceuticals·D J Buchsbaum, D T Curiel
Oct 9, 2007·Nuclear Medicine and Biology·Inna SerganovaRonald Blasberg
Dec 7, 2006·Neuroimaging Clinics of North America·Dawid Schellingerhout
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Oct 26, 2005·Nuclear Medicine and Biology·Inna Serganova, Ronald Blasberg
Nov 12, 2016·Cancer Gene Therapy·Z SaadatpourH Mirzaei
Apr 20, 2004·Molecular Therapy : the Journal of the American Society of Gene Therapy·Kenneth N BartonStephen L Brown
Mar 8, 2003·Technology in Cancer Research & Treatment·A H JacobsW D Heiss
May 14, 2004·Gene Therapy·K ShahR Weissleder
Oct 19, 2001·Oncogene·M D Schaller
Sep 25, 2001·Radiology·A MooreR Weissleder
Apr 5, 2014·Pest Management Science·Shiv S Kaundun
Jan 24, 2003·European Journal of Nuclear Medicine and Molecular Imaging·Norbert Avril, Frank M Bengel
Apr 12, 2003·Technology in Cancer Research & Treatment·Tandra R ChaudhuriKurt R Zinn
Nov 20, 2016·Cancer Gene Therapy·Z SaadatpourM Keshavarzi
Sep 8, 2001·Molecular Therapy : the Journal of the American Society of Gene Therapy·A HemminkiR D Alvarez

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