In vivo characterization of a reporter gene system for imaging hypoxia-induced gene expression

Sean Carlin, Andrei Pugachev, Xiaorong Sun, Sean Burke, Filip Claus, Joseph O'Donoghue, C. Clifton Ling, John L. Humm

Research output: Contribution to journalArticle

14 Citations (Scopus)

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 18F-fluoromisonidazole (18F-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 124I-2′-fluoro-2′-deoxy-1-β-d-arabinofuranosyl-5-iodouracil (124I-FIAU) 3 h prior to sacrifice. Statistical analysis of the spatial association between 124I-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 18F-FMISO. Two rats, each bearing a single R3327-AT/HRE tumor, were injected with 124I-FIAU (3 h before sacrifice) and 18F-FMISO (2 h before sacrifice). Statistical analysis of the spatial association between 18F-FMISO and 124I-FIAU on a pixel-by-pixel basis was performed. Results: Correlation coefficients between 124I-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 18F-FMISO and 124I-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.

Original languageEnglish (US)
Pages (from-to)821-831
Number of pages11
JournalNuclear Medicine and Biology
Volume36
Issue number7
DOIs
StatePublished - Oct 1 2009
Externally publishedYes

Fingerprint

Reporter Genes
Gene Expression
Neoplasms
Spatial Analysis
Staining and Labeling
Hypoxia
Gene Fusion
fialuridine
Prostatic Neoplasms
fluoromisonidazole
Cell Line
pimonidazole

Keywords

  • Cancer
  • Hypoxia
  • Nuclear medicine
  • PET tracer validation
  • Reporter gene

ASJC Scopus subject areas

  • Molecular Medicine
  • Radiology Nuclear Medicine and imaging
  • Cancer Research

Cite this

In vivo characterization of a reporter gene system for imaging hypoxia-induced gene expression. / Carlin, Sean; Pugachev, Andrei; Sun, Xiaorong; Burke, Sean; Claus, Filip; O'Donoghue, Joseph; Ling, C. Clifton; Humm, John L.

In: Nuclear Medicine and Biology, Vol. 36, No. 7, 01.10.2009, p. 821-831.

Research output: Contribution to journalArticle

Carlin, Sean ; Pugachev, Andrei ; Sun, Xiaorong ; Burke, Sean ; Claus, Filip ; O'Donoghue, Joseph ; Ling, C. Clifton ; Humm, John L. / In vivo characterization of a reporter gene system for imaging hypoxia-induced gene expression. In: Nuclear Medicine and Biology. 2009 ; Vol. 36, No. 7. pp. 821-831.
@article{c23bd0566cc445fc954570f6e0db86fe,
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 18F-fluoromisonidazole (18F-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 124I-2′-fluoro-2′-deoxy-1-β-d-arabinofuranosyl-5-iodouracil (124I-FIAU) 3 h prior to sacrifice. Statistical analysis of the spatial association between 124I-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 18F-FMISO. Two rats, each bearing a single R3327-AT/HRE tumor, were injected with 124I-FIAU (3 h before sacrifice) and 18F-FMISO (2 h before sacrifice). Statistical analysis of the spatial association between 18F-FMISO and 124I-FIAU on a pixel-by-pixel basis was performed. Results: Correlation coefficients between 124I-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 18F-FMISO and 124I-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.",
keywords = "Cancer, Hypoxia, Nuclear medicine, PET tracer validation, Reporter gene",
author = "Sean Carlin and Andrei Pugachev and Xiaorong Sun and Sean Burke and Filip Claus and Joseph O'Donoghue and Ling, {C. Clifton} and Humm, {John L.}",
year = "2009",
month = "10",
day = "1",
doi = "10.1016/j.nucmedbio.2009.06.006",
language = "English (US)",
volume = "36",
pages = "821--831",
journal = "International journal of radiation applications and instrumentation. Part B, Nuclear medicine and biology",
issn = "0969-8051",
publisher = "Elsevier Inc.",
number = "7",

}

TY - JOUR

T1 - In vivo characterization of a reporter gene system for imaging hypoxia-induced gene expression

AU - Carlin, Sean

AU - Pugachev, Andrei

AU - Sun, Xiaorong

AU - Burke, Sean

AU - Claus, Filip

AU - O'Donoghue, Joseph

AU - Ling, C. Clifton

AU - Humm, John L.

PY - 2009/10/1

Y1 - 2009/10/1

N2 - 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 18F-fluoromisonidazole (18F-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 124I-2′-fluoro-2′-deoxy-1-β-d-arabinofuranosyl-5-iodouracil (124I-FIAU) 3 h prior to sacrifice. Statistical analysis of the spatial association between 124I-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 18F-FMISO. Two rats, each bearing a single R3327-AT/HRE tumor, were injected with 124I-FIAU (3 h before sacrifice) and 18F-FMISO (2 h before sacrifice). Statistical analysis of the spatial association between 18F-FMISO and 124I-FIAU on a pixel-by-pixel basis was performed. Results: Correlation coefficients between 124I-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 18F-FMISO and 124I-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.

AB - 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 18F-fluoromisonidazole (18F-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 124I-2′-fluoro-2′-deoxy-1-β-d-arabinofuranosyl-5-iodouracil (124I-FIAU) 3 h prior to sacrifice. Statistical analysis of the spatial association between 124I-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 18F-FMISO. Two rats, each bearing a single R3327-AT/HRE tumor, were injected with 124I-FIAU (3 h before sacrifice) and 18F-FMISO (2 h before sacrifice). Statistical analysis of the spatial association between 18F-FMISO and 124I-FIAU on a pixel-by-pixel basis was performed. Results: Correlation coefficients between 124I-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 18F-FMISO and 124I-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.

KW - Cancer

KW - Hypoxia

KW - Nuclear medicine

KW - PET tracer validation

KW - Reporter gene

UR - http://www.scopus.com/inward/record.url?scp=69249215186&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=69249215186&partnerID=8YFLogxK

U2 - 10.1016/j.nucmedbio.2009.06.006

DO - 10.1016/j.nucmedbio.2009.06.006

M3 - Article

C2 - 19720294

AN - SCOPUS:69249215186

VL - 36

SP - 821

EP - 831

JO - International journal of radiation applications and instrumentation. Part B, Nuclear medicine and biology

JF - International journal of radiation applications and instrumentation. Part B, Nuclear medicine and biology

SN - 0969-8051

IS - 7

ER -