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A probasin promoter, conditionally replicating adenovirus that expresses the sodium iodide symporter (NIS) for radiovirotherapy of prostate cancer.

Trujillo MA, Oneal MJ, McDonough S, Qin R, Morris JC - Gene Ther. (2010)

Bottom Line: Conversely, the prostate cancer cell line PC-3 (androgen receptor negative) and the pancreatic cancer cell line Panc-1 were refractory to the viral cytopathic effect and did not support viral replication.Radioiodine uptake was readily measurable in LnCaP cells infected with Ad5PB_RSV-NIS 24 h post-infection, confirming NIS expression.Administration of therapeutic ¹³¹I after virus injection significantly increased survival probability in mice carrying xenografted LnCaP tumors compared with virotherapy alone.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, Division of Endocrinology, Diabetes, Metabolism, Nutrition, Mayo Clinic, Rochester, MN 55905, USA.

ABSTRACT
The sodium iodide symporter (NIS) directs the uptake and concentration of iodide in thyroid cells. We have extended the use of NIS-mediated radioiodine therapy to other types of cancer, we transferred and expressed the NIS gene into prostate, colon and breast cancer cells using adenoviral vectors. To improve vector efficiency we have developed a conditionally replicating adenovirus (CRAd) in which the E1a gene is driven by the prostate-specific promoter, Probasin and the cassette RSV promoter human NIScDNA-bGH polyA replaces the E3 region (CRAd Ad5PB_RSV-NIS). In vitro infection of the prostate cancer cell line LnCaP resulted in virus replication, cytolysis and release of infective viral particles. Conversely, the prostate cancer cell line PC-3 (androgen receptor negative) and the pancreatic cancer cell line Panc-1 were refractory to the viral cytopathic effect and did not support viral replication. Radioiodine uptake was readily measurable in LnCaP cells infected with Ad5PB_RSV-NIS 24 h post-infection, confirming NIS expression. In vivo, LnCaP tumor xenografts in nude-mice injected intratumorally with Ad5PB_RSV_NIS CRAd expressed NIS actively as evidenced by ⁹⁹Tc uptake and imaging. Administration of therapeutic ¹³¹I after virus injection significantly increased survival probability in mice carrying xenografted LnCaP tumors compared with virotherapy alone. These data indicate that Ad5PB_RSV_NIS replication is stringently restricted to androgen-positive prostate cancer cells and results in effective NIS expression and uptake of radioiodine. This construct may allow multimodal therapy, combining cytolytic virotherapy with radioiodine treatment, to be developed as a novel treatment for prostate cancer.

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Virus Mediated Cell KillingA) Cytopathic effect. Ad5PB_RSV-NIS-induced cytopathic effect was assessed by light microscopy. B) Virus-mediated cell-specific killing. The prostate cancer cell line LnCaP and the pancreatic cell line Panc-1 were infected with Ad5PB_RSV-NIS at the indicated MOI. At time points cell viability was assayed by MTS.
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Figure 1: Virus Mediated Cell KillingA) Cytopathic effect. Ad5PB_RSV-NIS-induced cytopathic effect was assessed by light microscopy. B) Virus-mediated cell-specific killing. The prostate cancer cell line LnCaP and the pancreatic cell line Panc-1 were infected with Ad5PB_RSV-NIS at the indicated MOI. At time points cell viability was assayed by MTS.

Mentions: We assayed the specificity of Ad5PB_RSV-NIS replication using several tests. We first examined the specificity of viral protein expression by Western blot for Probasin-driven E1A and Hexon expression (Not Shown). We next measure the specificity of cytopathic effect (CPE) by light microscopy. Cells were infected in triplicates with Ad5PB_RSV-NIS at MOI 1 and examined four days later for signs of CPE. The morphology of the PC-3 cells was unchanged after infection with Ad5PB_RSV-NIS. However, clear signs of CPE in LnCaP were detected under the same conditions (Figure 1A).


A probasin promoter, conditionally replicating adenovirus that expresses the sodium iodide symporter (NIS) for radiovirotherapy of prostate cancer.

Trujillo MA, Oneal MJ, McDonough S, Qin R, Morris JC - Gene Ther. (2010)

Virus Mediated Cell KillingA) Cytopathic effect. Ad5PB_RSV-NIS-induced cytopathic effect was assessed by light microscopy. B) Virus-mediated cell-specific killing. The prostate cancer cell line LnCaP and the pancreatic cell line Panc-1 were infected with Ad5PB_RSV-NIS at the indicated MOI. At time points cell viability was assayed by MTS.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2914818&req=5

Figure 1: Virus Mediated Cell KillingA) Cytopathic effect. Ad5PB_RSV-NIS-induced cytopathic effect was assessed by light microscopy. B) Virus-mediated cell-specific killing. The prostate cancer cell line LnCaP and the pancreatic cell line Panc-1 were infected with Ad5PB_RSV-NIS at the indicated MOI. At time points cell viability was assayed by MTS.
Mentions: We assayed the specificity of Ad5PB_RSV-NIS replication using several tests. We first examined the specificity of viral protein expression by Western blot for Probasin-driven E1A and Hexon expression (Not Shown). We next measure the specificity of cytopathic effect (CPE) by light microscopy. Cells were infected in triplicates with Ad5PB_RSV-NIS at MOI 1 and examined four days later for signs of CPE. The morphology of the PC-3 cells was unchanged after infection with Ad5PB_RSV-NIS. However, clear signs of CPE in LnCaP were detected under the same conditions (Figure 1A).

Bottom Line: Conversely, the prostate cancer cell line PC-3 (androgen receptor negative) and the pancreatic cancer cell line Panc-1 were refractory to the viral cytopathic effect and did not support viral replication.Radioiodine uptake was readily measurable in LnCaP cells infected with Ad5PB_RSV-NIS 24 h post-infection, confirming NIS expression.Administration of therapeutic ¹³¹I after virus injection significantly increased survival probability in mice carrying xenografted LnCaP tumors compared with virotherapy alone.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, Division of Endocrinology, Diabetes, Metabolism, Nutrition, Mayo Clinic, Rochester, MN 55905, USA.

ABSTRACT
The sodium iodide symporter (NIS) directs the uptake and concentration of iodide in thyroid cells. We have extended the use of NIS-mediated radioiodine therapy to other types of cancer, we transferred and expressed the NIS gene into prostate, colon and breast cancer cells using adenoviral vectors. To improve vector efficiency we have developed a conditionally replicating adenovirus (CRAd) in which the E1a gene is driven by the prostate-specific promoter, Probasin and the cassette RSV promoter human NIScDNA-bGH polyA replaces the E3 region (CRAd Ad5PB_RSV-NIS). In vitro infection of the prostate cancer cell line LnCaP resulted in virus replication, cytolysis and release of infective viral particles. Conversely, the prostate cancer cell line PC-3 (androgen receptor negative) and the pancreatic cancer cell line Panc-1 were refractory to the viral cytopathic effect and did not support viral replication. Radioiodine uptake was readily measurable in LnCaP cells infected with Ad5PB_RSV-NIS 24 h post-infection, confirming NIS expression. In vivo, LnCaP tumor xenografts in nude-mice injected intratumorally with Ad5PB_RSV_NIS CRAd expressed NIS actively as evidenced by ⁹⁹Tc uptake and imaging. Administration of therapeutic ¹³¹I after virus injection significantly increased survival probability in mice carrying xenografted LnCaP tumors compared with virotherapy alone. These data indicate that Ad5PB_RSV_NIS replication is stringently restricted to androgen-positive prostate cancer cells and results in effective NIS expression and uptake of radioiodine. This construct may allow multimodal therapy, combining cytolytic virotherapy with radioiodine treatment, to be developed as a novel treatment for prostate cancer.

Show MeSH
Related in: MedlinePlus