Imaging of HIF-1-active tumor hypoxia using a protein effectively delivered to and specifically stabilized in HIF-1-active tumor cells

Takashi Kudo, Masashi Ueda, Yuji Kuge, Takahiro Mukai, Shotaro Tanaka, Maki Masutani, Yasushi Kiyono, Shinae Kizaka-Kondoh, Masahiro Hiraoka, Hideo Saji

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Hypoxia-inducible factor-1 (HIF-1) plays an important role in malignant tumor progression and in the development of resistance to radiotherapy. We designed a novel fusion protein (PTD-ODD-SAV [POS]) consisting of a protein transduction domain (PTD), streptavidin (SAV), and a portion of the oxygen-dependent degradation domain (ODD) of HIF-1α that confers the same oxygen-dependent regulation as HIF-1α on POS. (3- 123/125Iiodobenzoyl) norbiotinamide (123/125I-IBB) was conjugated to the SAV moiety of POS to synthesize 123/125I-IBB- labeled POS (123/125I-IPOS). The purpose of this study was to evaluate the feasibility of 123I-IPOS as an imaging probe for HIF-1-active tumor hypoxia. Methods: After a 24-h incubation of 125I-IPOS with various tumor cell lines under either normoxic (20% O2) or hypoxic (0.1% O2) conditions, the intracellular radioactivity was investigated. Then, the biodistribution of 123/125I-IPOS was examined with tumor-implanted mice, and an in vivo imaging study was performed. The tumoral accumulation of 125I-IPOS was compared with HIF-1 activity using the mice carrying tumors with the HIF-1-dependent luciferase reporter gene. Furthermore, the intratumoral localization of 125I-IPOS was examined by the autoradiographic study, and then the same slide was subjected to immunostaining for pimonidazole, which is the hypoxic marker. Results: The ratios of radioactivity in hypoxic cells to that in normoxic cells were more than 2. These results indicate incorporation of 125I-IPOS into these cells and degradation of 125I-IPOS by normoxic tumor cells. In the biodistribution study, 125I-IPOS accumulated in the tumor (1.4 ± 0.3 percentage injected dose per gram) 24 h after administration. At that time, 125I-IPOS showed high tumor-to-blood and tumor-to-muscle ratios (5.1 ± 0.3 and 14.0 ± 3.9, respectively). The tumors were clearly visualized by in vivo imaging 24 h after 123I-IPOS injection (tumor-to-muscle ratio was 9.6). The tumoral accumulation of 125I-IPOS correlated with HIF-1 activity (R = 0.71, P <0.05), and its intratumoral distribution coincided with the hypoxic regions. Conclusion: 123I-IPOS is a potential probe for the imaging of HIF-1 activity in tumors. Given the role of HIF-1 in tumor biology, its detection may be considered an indicator of aggressive cancer phenotypes.

Original languageEnglish
Pages (from-to)942-949
Number of pages8
JournalJournal of Nuclear Medicine
Volume50
Issue number6
DOIs
Publication statusPublished - Jun 1 2009
Externally publishedYes

Fingerprint

Hypoxia-Inducible Factor 1
Neoplasms
Proteins
Streptavidin
Oxygen
Radioactivity
Tumor Hypoxia
Muscles
Tumor Cell Line
Luciferases
Reporter Genes
Radiotherapy

Keywords

  • Hypoxia-inducible factor-1 (HIF-1)
  • Molecular imaging
  • Oncology
  • Oxygen-dependent degradation (ODD)
  • Protein transduction domain (PTD)
  • Radiopharmaceuticals
  • Tumor hypoxia

ASJC Scopus subject areas

  • Radiology Nuclear Medicine and imaging

Cite this

Imaging of HIF-1-active tumor hypoxia using a protein effectively delivered to and specifically stabilized in HIF-1-active tumor cells. / Kudo, Takashi; Ueda, Masashi; Kuge, Yuji; Mukai, Takahiro; Tanaka, Shotaro; Masutani, Maki; Kiyono, Yasushi; Kizaka-Kondoh, Shinae; Hiraoka, Masahiro; Saji, Hideo.

In: Journal of Nuclear Medicine, Vol. 50, No. 6, 01.06.2009, p. 942-949.

Research output: Contribution to journalArticle

Kudo, T, Ueda, M, Kuge, Y, Mukai, T, Tanaka, S, Masutani, M, Kiyono, Y, Kizaka-Kondoh, S, Hiraoka, M & Saji, H 2009, 'Imaging of HIF-1-active tumor hypoxia using a protein effectively delivered to and specifically stabilized in HIF-1-active tumor cells', Journal of Nuclear Medicine, vol. 50, no. 6, pp. 942-949. https://doi.org/10.2967/jnumed.108.061119
Kudo, Takashi ; Ueda, Masashi ; Kuge, Yuji ; Mukai, Takahiro ; Tanaka, Shotaro ; Masutani, Maki ; Kiyono, Yasushi ; Kizaka-Kondoh, Shinae ; Hiraoka, Masahiro ; Saji, Hideo. / Imaging of HIF-1-active tumor hypoxia using a protein effectively delivered to and specifically stabilized in HIF-1-active tumor cells. In: Journal of Nuclear Medicine. 2009 ; Vol. 50, No. 6. pp. 942-949.
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abstract = "Hypoxia-inducible factor-1 (HIF-1) plays an important role in malignant tumor progression and in the development of resistance to radiotherapy. We designed a novel fusion protein (PTD-ODD-SAV [POS]) consisting of a protein transduction domain (PTD), streptavidin (SAV), and a portion of the oxygen-dependent degradation domain (ODD) of HIF-1α that confers the same oxygen-dependent regulation as HIF-1α on POS. (3- 123/125Iiodobenzoyl) norbiotinamide (123/125I-IBB) was conjugated to the SAV moiety of POS to synthesize 123/125I-IBB- labeled POS (123/125I-IPOS). The purpose of this study was to evaluate the feasibility of 123I-IPOS as an imaging probe for HIF-1-active tumor hypoxia. Methods: After a 24-h incubation of 125I-IPOS with various tumor cell lines under either normoxic (20{\%} O2) or hypoxic (0.1{\%} O2) conditions, the intracellular radioactivity was investigated. Then, the biodistribution of 123/125I-IPOS was examined with tumor-implanted mice, and an in vivo imaging study was performed. The tumoral accumulation of 125I-IPOS was compared with HIF-1 activity using the mice carrying tumors with the HIF-1-dependent luciferase reporter gene. Furthermore, the intratumoral localization of 125I-IPOS was examined by the autoradiographic study, and then the same slide was subjected to immunostaining for pimonidazole, which is the hypoxic marker. Results: The ratios of radioactivity in hypoxic cells to that in normoxic cells were more than 2. These results indicate incorporation of 125I-IPOS into these cells and degradation of 125I-IPOS by normoxic tumor cells. In the biodistribution study, 125I-IPOS accumulated in the tumor (1.4 ± 0.3 percentage injected dose per gram) 24 h after administration. At that time, 125I-IPOS showed high tumor-to-blood and tumor-to-muscle ratios (5.1 ± 0.3 and 14.0 ± 3.9, respectively). The tumors were clearly visualized by in vivo imaging 24 h after 123I-IPOS injection (tumor-to-muscle ratio was 9.6). The tumoral accumulation of 125I-IPOS correlated with HIF-1 activity (R = 0.71, P <0.05), and its intratumoral distribution coincided with the hypoxic regions. Conclusion: 123I-IPOS is a potential probe for the imaging of HIF-1 activity in tumors. Given the role of HIF-1 in tumor biology, its detection may be considered an indicator of aggressive cancer phenotypes.",
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TY - JOUR

T1 - Imaging of HIF-1-active tumor hypoxia using a protein effectively delivered to and specifically stabilized in HIF-1-active tumor cells

AU - Kudo, Takashi

AU - Ueda, Masashi

AU - Kuge, Yuji

AU - Mukai, Takahiro

AU - Tanaka, Shotaro

AU - Masutani, Maki

AU - Kiyono, Yasushi

AU - Kizaka-Kondoh, Shinae

AU - Hiraoka, Masahiro

AU - Saji, Hideo

PY - 2009/6/1

Y1 - 2009/6/1

N2 - Hypoxia-inducible factor-1 (HIF-1) plays an important role in malignant tumor progression and in the development of resistance to radiotherapy. We designed a novel fusion protein (PTD-ODD-SAV [POS]) consisting of a protein transduction domain (PTD), streptavidin (SAV), and a portion of the oxygen-dependent degradation domain (ODD) of HIF-1α that confers the same oxygen-dependent regulation as HIF-1α on POS. (3- 123/125Iiodobenzoyl) norbiotinamide (123/125I-IBB) was conjugated to the SAV moiety of POS to synthesize 123/125I-IBB- labeled POS (123/125I-IPOS). The purpose of this study was to evaluate the feasibility of 123I-IPOS as an imaging probe for HIF-1-active tumor hypoxia. Methods: After a 24-h incubation of 125I-IPOS with various tumor cell lines under either normoxic (20% O2) or hypoxic (0.1% O2) conditions, the intracellular radioactivity was investigated. Then, the biodistribution of 123/125I-IPOS was examined with tumor-implanted mice, and an in vivo imaging study was performed. The tumoral accumulation of 125I-IPOS was compared with HIF-1 activity using the mice carrying tumors with the HIF-1-dependent luciferase reporter gene. Furthermore, the intratumoral localization of 125I-IPOS was examined by the autoradiographic study, and then the same slide was subjected to immunostaining for pimonidazole, which is the hypoxic marker. Results: The ratios of radioactivity in hypoxic cells to that in normoxic cells were more than 2. These results indicate incorporation of 125I-IPOS into these cells and degradation of 125I-IPOS by normoxic tumor cells. In the biodistribution study, 125I-IPOS accumulated in the tumor (1.4 ± 0.3 percentage injected dose per gram) 24 h after administration. At that time, 125I-IPOS showed high tumor-to-blood and tumor-to-muscle ratios (5.1 ± 0.3 and 14.0 ± 3.9, respectively). The tumors were clearly visualized by in vivo imaging 24 h after 123I-IPOS injection (tumor-to-muscle ratio was 9.6). The tumoral accumulation of 125I-IPOS correlated with HIF-1 activity (R = 0.71, P <0.05), and its intratumoral distribution coincided with the hypoxic regions. Conclusion: 123I-IPOS is a potential probe for the imaging of HIF-1 activity in tumors. Given the role of HIF-1 in tumor biology, its detection may be considered an indicator of aggressive cancer phenotypes.

AB - Hypoxia-inducible factor-1 (HIF-1) plays an important role in malignant tumor progression and in the development of resistance to radiotherapy. We designed a novel fusion protein (PTD-ODD-SAV [POS]) consisting of a protein transduction domain (PTD), streptavidin (SAV), and a portion of the oxygen-dependent degradation domain (ODD) of HIF-1α that confers the same oxygen-dependent regulation as HIF-1α on POS. (3- 123/125Iiodobenzoyl) norbiotinamide (123/125I-IBB) was conjugated to the SAV moiety of POS to synthesize 123/125I-IBB- labeled POS (123/125I-IPOS). The purpose of this study was to evaluate the feasibility of 123I-IPOS as an imaging probe for HIF-1-active tumor hypoxia. Methods: After a 24-h incubation of 125I-IPOS with various tumor cell lines under either normoxic (20% O2) or hypoxic (0.1% O2) conditions, the intracellular radioactivity was investigated. Then, the biodistribution of 123/125I-IPOS was examined with tumor-implanted mice, and an in vivo imaging study was performed. The tumoral accumulation of 125I-IPOS was compared with HIF-1 activity using the mice carrying tumors with the HIF-1-dependent luciferase reporter gene. Furthermore, the intratumoral localization of 125I-IPOS was examined by the autoradiographic study, and then the same slide was subjected to immunostaining for pimonidazole, which is the hypoxic marker. Results: The ratios of radioactivity in hypoxic cells to that in normoxic cells were more than 2. These results indicate incorporation of 125I-IPOS into these cells and degradation of 125I-IPOS by normoxic tumor cells. In the biodistribution study, 125I-IPOS accumulated in the tumor (1.4 ± 0.3 percentage injected dose per gram) 24 h after administration. At that time, 125I-IPOS showed high tumor-to-blood and tumor-to-muscle ratios (5.1 ± 0.3 and 14.0 ± 3.9, respectively). The tumors were clearly visualized by in vivo imaging 24 h after 123I-IPOS injection (tumor-to-muscle ratio was 9.6). The tumoral accumulation of 125I-IPOS correlated with HIF-1 activity (R = 0.71, P <0.05), and its intratumoral distribution coincided with the hypoxic regions. Conclusion: 123I-IPOS is a potential probe for the imaging of HIF-1 activity in tumors. Given the role of HIF-1 in tumor biology, its detection may be considered an indicator of aggressive cancer phenotypes.

KW - Hypoxia-inducible factor-1 (HIF-1)

KW - Molecular imaging

KW - Oncology

KW - Oxygen-dependent degradation (ODD)

KW - Protein transduction domain (PTD)

KW - Radiopharmaceuticals

KW - Tumor hypoxia

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