In vivo characterization of a reporter gene system for imaging hypoxia-induced gene expression Journal Article
| Authors: | Carlin, S.; Pugachev, A.; Sun, X.; Burke, S.; Claus, F.; O'Donoghue, J.; Ling, C. C.; Humm, J. L. |
| Article Title: | In vivo characterization of a reporter gene system for imaging hypoxia-induced gene expression |
| Abstract: | Purpose: To characterize a tumor model containing a hypoxia-inducible reporter gene and to demonstrate utility by comparison of reporter gene expression to the uptake and distribution of the hypoxia tracer <sup>18</sup>F-fluoromisonidazole (<sup>18</sup>F-FMISO). Methods: Three tumors derived from the rat prostate cancer cell line R3327-AT were grown in each of two rats as follows: (1) parental R3327-AT, (2) positive control R3327-AT/PC in which the HSV1-tkeGFP fusion reporter gene was expressed constitutively, (3) R3327-AT/HRE in which the reporter gene was placed under the control of a hypoxia-inducible factor-responsive promoter sequence (HRE). Animals were coadministered a hypoxia-specific marker (pimonidazole) and the reporter gene probe <sup>124</sup>I-2′-fluoro-2′-deoxy-1-β-d-arabinofuranosyl-5-iodouracil (<sup>124</sup>I-FIAU) 3 h prior to sacrifice. Statistical analysis of the spatial association between <sup>124</sup>I-FIAU uptake and pimonidazole fluorescent staining intensity was then performed on a pixel-by-pixel basis. Utility of this system was demonstrated by assessment of reporter gene expression versus the exogenous hypoxia probe <sup>18</sup>F-FMISO. Two rats, each bearing a single R3327-AT/HRE tumor, were injected with <sup>124</sup>I-FIAU (3 h before sacrifice) and <sup>18</sup>F-FMISO (2 h before sacrifice). Statistical analysis of the spatial association between <sup>18</sup>F-FMISO and <sup>124</sup>I-FIAU on a pixel-by-pixel basis was performed. Results: Correlation coefficients between <sup>124</sup>I-FIAU uptake and pimonidazole staining intensity were: 0.11 in R3327-AT tumors, -0.66 in R3327-AT/PC and 0.76 in R3327-AT/HRE, confirming that only in the R3327-AT/HRE tumor was HSV1-tkeGFP gene expression associated with hypoxia. Correlation coefficients between <sup>18</sup>F-FMISO and <sup>124</sup>I-FIAU uptakes in R3327-AT/HRE tumors were r=0.56, demonstrating good spatial correspondence between the two tracers. Conclusions: We have confirmed hypoxia-specific expression of the HSV1-tkeGFP fusion gene in the R3327-AT/HRE tumor model and demonstrated the utility of this model for the evaluation of radiolabeled hypoxia tracers. © 2009 Elsevier Inc. All rights reserved. |
| Keywords: | controlled study; unclassified drug; promoter region; nonhuman; animals; animal tissue; gene expression; fluorescence; green fluorescent protein; animal experiment; in vivo study; cancer cell culture; cell line, tumor; prostate cancer; hypoxia; gene expression regulation, neoplastic; correlation coefficient; statistical analysis; drug distribution; drug uptake; isotope labeling; tumor burden; rat; fusion gene; radiopharmaceutical agent; reporter gene; imaging; nuclear medicine; pet tracer validation; 1 fluoro 3 (2 nitro 1 imidazolyl) 2 propanol f 18; fialuridine; fialuridine i 124; hoe 33342; hypoxia inducible factor; iodine 124; pimonidazole; thymidine kinase; gene probe; tumor model; cell hypoxia; genes, reporter; herpesvirus 1, human; radioactive tracers; rats; response elements |
| Journal Title: | Nuclear Medicine and Biology |
| Volume: | 36 |
| Issue: | 7 |
| ISSN: | 0969-8051 |
| Publisher: | Elsevier Science Inc. |
| Date Published: | 2009-10-01 |
| Start Page: | 821 |
| End Page: | 831 |
| Language: | English |
| DOI: | 10.1016/j.nucmedbio.2009.06.006 |
| PUBMED: | 19720294 |
| PROVIDER: | scopus |
| PMCID: | PMC2754273 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 1" - "Export Date: 30 November 2010" - "CODEN: NMBIE" - "Source: Scopus" |
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