Utilizing tumor hypoxia to enhance oncolytic viral therapy in colorectal metastases

Maura Reinblatt, Richard H. Pin, Howard J. Federoff, Yuman Fong, B. Mark Evers, Kevin E. Behrns, W. Roy Smythe, William C. Lineaweaver, Max Langham

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Objective: To determine the effects of hypoxia-induced ribonucleotide reductase (RR) production on herpes oncolytic viral therapy. Summary Background Data: Hypoxia is a common tumor condition correlated with therapeutic resistance and metastases. Attenuated viruses offer a unique cancer treatment by specifically infecting and lysing tumor cells. G207 is an oncolytic herpes virus deficient in RR, a rate-limiting enzyme for viral replication. Methods: A multimerized hypoxia-responsive enhancer was constructed (10xHRE) and functionally tested by luciferase assay. 10xHRE was cloned upstream of UL39, the gene encoding the large subunit of RR (10xHRE-UL39). CT26 murine colorectal cancer cells were transfected with 10xHRE-UL39, incubated in hypoxia (1% O 2) or normoxia (21% O2), and infected with G207 for cytotoxicity assays. CT26 liver metastases, with or without 10xHRE-UL39, were created in syngeneic Balb/C mice (n = 40). Livers were treated with G207 or saline. Tumors were assessed and stained immunohistochemically for G207. Results: 10xHRE increased luciferase expression 33-fold in hypoxia versus controls (P < 0.001). In normoxia, 10xHRE-UL39 transfection did not improve G207 cytotoxicity. In hypoxia, G207 cytotoxicity increased 87% with 10xHRE-UL39 transfection versus nontransfected cells (P < 0.001). CT26 were resistant to G207 alone. Combining 10xHRE-UL39 with G207 resulted in a 66% decrease in tumor weights (P < 0.0001) and a 65% reduction in tumor nodules (P < 0.0001) versus G207 monotherapy. 10xHRE-UL39-transfected tumors demonstrated greater viral staining. Conclusions: Hypoxia-driven RR production significantly enhances viral cytotoxicity in vitro and reduces tumor burden in vivo. G207 combined with RR under hypoxic control is a promising treatment for colorectal cancer, which would otherwise be resistant to oncolytic herpes virus alone.

Original languageEnglish (US)
Pages (from-to)892-902
Number of pages11
JournalAnnals of surgery
Volume239
Issue number6
DOIs
StatePublished - Jun 1 2004

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Ribonucleotide Reductases
Neoplasm Metastasis
Oncolytic Viruses
Neoplasms
Tumor Burden
Luciferases
Therapeutics
Transfection
Colorectal Neoplasms
Liver
Tumor Hypoxia
Hypoxia
Staining and Labeling
Viruses
Enzymes
Genes

All Science Journal Classification (ASJC) codes

  • Surgery

Cite this

Utilizing tumor hypoxia to enhance oncolytic viral therapy in colorectal metastases. / Reinblatt, Maura; Pin, Richard H.; Federoff, Howard J.; Fong, Yuman; Evers, B. Mark; Behrns, Kevin E.; Smythe, W. Roy; Lineaweaver, William C.; Langham, Max.

In: Annals of surgery, Vol. 239, No. 6, 01.06.2004, p. 892-902.

Research output: Contribution to journalArticle

Reinblatt, M, Pin, RH, Federoff, HJ, Fong, Y, Evers, BM, Behrns, KE, Smythe, WR, Lineaweaver, WC & Langham, M 2004, 'Utilizing tumor hypoxia to enhance oncolytic viral therapy in colorectal metastases', Annals of surgery, vol. 239, no. 6, pp. 892-902. https://doi.org/10.1097/01.sla.0000128308.36393.38
Reinblatt M, Pin RH, Federoff HJ, Fong Y, Evers BM, Behrns KE et al. Utilizing tumor hypoxia to enhance oncolytic viral therapy in colorectal metastases. Annals of surgery. 2004 Jun 1;239(6):892-902. https://doi.org/10.1097/01.sla.0000128308.36393.38
Reinblatt, Maura ; Pin, Richard H. ; Federoff, Howard J. ; Fong, Yuman ; Evers, B. Mark ; Behrns, Kevin E. ; Smythe, W. Roy ; Lineaweaver, William C. ; Langham, Max. / Utilizing tumor hypoxia to enhance oncolytic viral therapy in colorectal metastases. In: Annals of surgery. 2004 ; Vol. 239, No. 6. pp. 892-902.
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abstract = "Objective: To determine the effects of hypoxia-induced ribonucleotide reductase (RR) production on herpes oncolytic viral therapy. Summary Background Data: Hypoxia is a common tumor condition correlated with therapeutic resistance and metastases. Attenuated viruses offer a unique cancer treatment by specifically infecting and lysing tumor cells. G207 is an oncolytic herpes virus deficient in RR, a rate-limiting enzyme for viral replication. Methods: A multimerized hypoxia-responsive enhancer was constructed (10xHRE) and functionally tested by luciferase assay. 10xHRE was cloned upstream of UL39, the gene encoding the large subunit of RR (10xHRE-UL39). CT26 murine colorectal cancer cells were transfected with 10xHRE-UL39, incubated in hypoxia (1{\%} O 2) or normoxia (21{\%} O2), and infected with G207 for cytotoxicity assays. CT26 liver metastases, with or without 10xHRE-UL39, were created in syngeneic Balb/C mice (n = 40). Livers were treated with G207 or saline. Tumors were assessed and stained immunohistochemically for G207. Results: 10xHRE increased luciferase expression 33-fold in hypoxia versus controls (P < 0.001). In normoxia, 10xHRE-UL39 transfection did not improve G207 cytotoxicity. In hypoxia, G207 cytotoxicity increased 87{\%} with 10xHRE-UL39 transfection versus nontransfected cells (P < 0.001). CT26 were resistant to G207 alone. Combining 10xHRE-UL39 with G207 resulted in a 66{\%} decrease in tumor weights (P < 0.0001) and a 65{\%} reduction in tumor nodules (P < 0.0001) versus G207 monotherapy. 10xHRE-UL39-transfected tumors demonstrated greater viral staining. Conclusions: Hypoxia-driven RR production significantly enhances viral cytotoxicity in vitro and reduces tumor burden in vivo. G207 combined with RR under hypoxic control is a promising treatment for colorectal cancer, which would otherwise be resistant to oncolytic herpes virus alone.",
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T1 - Utilizing tumor hypoxia to enhance oncolytic viral therapy in colorectal metastases

AU - Reinblatt, Maura

AU - Pin, Richard H.

AU - Federoff, Howard J.

AU - Fong, Yuman

AU - Evers, B. Mark

AU - Behrns, Kevin E.

AU - Smythe, W. Roy

AU - Lineaweaver, William C.

AU - Langham, Max

PY - 2004/6/1

Y1 - 2004/6/1

N2 - Objective: To determine the effects of hypoxia-induced ribonucleotide reductase (RR) production on herpes oncolytic viral therapy. Summary Background Data: Hypoxia is a common tumor condition correlated with therapeutic resistance and metastases. Attenuated viruses offer a unique cancer treatment by specifically infecting and lysing tumor cells. G207 is an oncolytic herpes virus deficient in RR, a rate-limiting enzyme for viral replication. Methods: A multimerized hypoxia-responsive enhancer was constructed (10xHRE) and functionally tested by luciferase assay. 10xHRE was cloned upstream of UL39, the gene encoding the large subunit of RR (10xHRE-UL39). CT26 murine colorectal cancer cells were transfected with 10xHRE-UL39, incubated in hypoxia (1% O 2) or normoxia (21% O2), and infected with G207 for cytotoxicity assays. CT26 liver metastases, with or without 10xHRE-UL39, were created in syngeneic Balb/C mice (n = 40). Livers were treated with G207 or saline. Tumors were assessed and stained immunohistochemically for G207. Results: 10xHRE increased luciferase expression 33-fold in hypoxia versus controls (P < 0.001). In normoxia, 10xHRE-UL39 transfection did not improve G207 cytotoxicity. In hypoxia, G207 cytotoxicity increased 87% with 10xHRE-UL39 transfection versus nontransfected cells (P < 0.001). CT26 were resistant to G207 alone. Combining 10xHRE-UL39 with G207 resulted in a 66% decrease in tumor weights (P < 0.0001) and a 65% reduction in tumor nodules (P < 0.0001) versus G207 monotherapy. 10xHRE-UL39-transfected tumors demonstrated greater viral staining. Conclusions: Hypoxia-driven RR production significantly enhances viral cytotoxicity in vitro and reduces tumor burden in vivo. G207 combined with RR under hypoxic control is a promising treatment for colorectal cancer, which would otherwise be resistant to oncolytic herpes virus alone.

AB - Objective: To determine the effects of hypoxia-induced ribonucleotide reductase (RR) production on herpes oncolytic viral therapy. Summary Background Data: Hypoxia is a common tumor condition correlated with therapeutic resistance and metastases. Attenuated viruses offer a unique cancer treatment by specifically infecting and lysing tumor cells. G207 is an oncolytic herpes virus deficient in RR, a rate-limiting enzyme for viral replication. Methods: A multimerized hypoxia-responsive enhancer was constructed (10xHRE) and functionally tested by luciferase assay. 10xHRE was cloned upstream of UL39, the gene encoding the large subunit of RR (10xHRE-UL39). CT26 murine colorectal cancer cells were transfected with 10xHRE-UL39, incubated in hypoxia (1% O 2) or normoxia (21% O2), and infected with G207 for cytotoxicity assays. CT26 liver metastases, with or without 10xHRE-UL39, were created in syngeneic Balb/C mice (n = 40). Livers were treated with G207 or saline. Tumors were assessed and stained immunohistochemically for G207. Results: 10xHRE increased luciferase expression 33-fold in hypoxia versus controls (P < 0.001). In normoxia, 10xHRE-UL39 transfection did not improve G207 cytotoxicity. In hypoxia, G207 cytotoxicity increased 87% with 10xHRE-UL39 transfection versus nontransfected cells (P < 0.001). CT26 were resistant to G207 alone. Combining 10xHRE-UL39 with G207 resulted in a 66% decrease in tumor weights (P < 0.0001) and a 65% reduction in tumor nodules (P < 0.0001) versus G207 monotherapy. 10xHRE-UL39-transfected tumors demonstrated greater viral staining. Conclusions: Hypoxia-driven RR production significantly enhances viral cytotoxicity in vitro and reduces tumor burden in vivo. G207 combined with RR under hypoxic control is a promising treatment for colorectal cancer, which would otherwise be resistant to oncolytic herpes virus alone.

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