Limits...
Prevention of adverse events of interferon γ gene therapy by gene delivery of interferon γ-heparin-binding domain fusion protein in mice.

Ando M, Takahashi Y, Yamashita T, Fujimoto M, Nishikawa M, Watanabe Y, Takakura Y - Mol Ther Methods Clin Dev (2014)

Bottom Line: The serum concentration of IFNγ-HBD2 and IFNγ-HBD3 after gene delivery was lower than that of IFNγ or IFNγ-HBD1.Gene delivery of IFNγ-HBD2, but not of IFNγ-HBD3, effectively increased the mRNA expression of IFNγ-inducible genes in the liver, suggesting liver-specific distribution of IFNγ-HBD2.Gene delivery of IFNγ-HBD2-suppressed tumor growth in the liver as efficiently as that of IFNγ with much less symptoms of adverse effects.

View Article: PubMed Central - PubMed

Affiliation: Department of Biopharmaceutics and Drug Metabolism , Graduate School of Pharmaceutical Sciences, Kyoto University , Sakyo-ku, Kyoto, Japan.

ABSTRACT
Sustained gene delivery of interferon (IFN) γ can be an effective treatment, but our previous study showed high levels of IFNγ-induced adverse events, including the loss of body weight. These unwanted events could be reduced by target-specific delivery of IFNγ after in vivo gene transfer. To achieve this, we selected the heparin-binding domain (HBD) of extracellular superoxide dismutase as a molecule to anchor IFNγ to the cell surface. We designed three IFNγ derivatives, IFNγ-HBD1, IFNγ-HBD2, and IFNγ-HBD3, each of which had 1, 2, or 3 HBDs, respectively. Each plasmid-encoding fusion proteins was delivered to the liver, a model target in this study, by hydrodynamic tail vein injection. The serum concentration of IFNγ-HBD2 and IFNγ-HBD3 after gene delivery was lower than that of IFNγ or IFNγ-HBD1. Gene delivery of IFNγ-HBD2, but not of IFNγ-HBD3, effectively increased the mRNA expression of IFNγ-inducible genes in the liver, suggesting liver-specific distribution of IFNγ-HBD2. Gene delivery of IFNγ-HBD2-suppressed tumor growth in the liver as efficiently as that of IFNγ with much less symptoms of adverse effects. These results indicate that the adverse events of IFNγ gene transfer can be prevented by gene delivery of IFNγ-HBD2, a fusion protein with high cell surface affinity.

No MeSH data available.


Related in: MedlinePlus

Dose-dependent IFNγ expression and IFNγ induced gene expression in mouse liver. ICR mice were injected with different doses of pCpG-IFNγ (solid circles), pCpG-IFNγ-HBD1 (open circles), pCpG-IFNγ-HBD2 (open triangles), and pCpG-IFNγ-HBD3 (open squares) by hydrodynamic gene transfer. Three days after gene transfer, mice were sacrificed and the livers were collected. (a) The mRNA expression of IFNγ, (b) The mRNA expression of SOCS1, (c) The mRNA expression of γ-IP10. Results are expressed as mean ± SEM (n = 3 or 4). HBD, heparin-binding domain; ICR, Institute of Cancer Research; IFN, interferon.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4362348&req=5

fig4: Dose-dependent IFNγ expression and IFNγ induced gene expression in mouse liver. ICR mice were injected with different doses of pCpG-IFNγ (solid circles), pCpG-IFNγ-HBD1 (open circles), pCpG-IFNγ-HBD2 (open triangles), and pCpG-IFNγ-HBD3 (open squares) by hydrodynamic gene transfer. Three days after gene transfer, mice were sacrificed and the livers were collected. (a) The mRNA expression of IFNγ, (b) The mRNA expression of SOCS1, (c) The mRNA expression of γ-IP10. Results are expressed as mean ± SEM (n = 3 or 4). HBD, heparin-binding domain; ICR, Institute of Cancer Research; IFN, interferon.

Mentions: pCpG vectors were used instead of pcDNA3 vectors for animal studies, because the former are more efficient in terms of transgene expression than the latter in mice. Mice received a hydrodynamic injection of any of the plasmids at different doses, and the livers were harvested 3 days after injection. Figure 4 shows the mRNA expression of IFNγ (or IFNγ-HBD fusion proteins), a suppressor of cytokine signaling 1 (SOCS1), and IFNγ inducible protein 10 (γ-IP10) in the mouse liver. The mRNA expression of the transgenes was dose dependent in all the groups and did not differ markedly from one to the other. The expression of IFNγ significantly increased the mRNA expression of SOCS1 and γ-IP10, indicating that IFNγ expressed in the liver is biologically active. The expression of these genes in the pCpG-IFNγ-HBD1–injected group was increased to a similar level in the pCpG-IFNγ–injected group. On the other hand, the expression in the pCpG-IFNγ-HBD2– and pCpG-IFNγ-HBD3–injected groups was significantly lower than that in the pCpG-IFNγ– or pCpG-IFNγ-HBD1–injected group, and the mRNA expression in the pCpG-IFNγ-HBD3–injected group was almost negligible over the dose range examined.


Prevention of adverse events of interferon γ gene therapy by gene delivery of interferon γ-heparin-binding domain fusion protein in mice.

Ando M, Takahashi Y, Yamashita T, Fujimoto M, Nishikawa M, Watanabe Y, Takakura Y - Mol Ther Methods Clin Dev (2014)

Dose-dependent IFNγ expression and IFNγ induced gene expression in mouse liver. ICR mice were injected with different doses of pCpG-IFNγ (solid circles), pCpG-IFNγ-HBD1 (open circles), pCpG-IFNγ-HBD2 (open triangles), and pCpG-IFNγ-HBD3 (open squares) by hydrodynamic gene transfer. Three days after gene transfer, mice were sacrificed and the livers were collected. (a) The mRNA expression of IFNγ, (b) The mRNA expression of SOCS1, (c) The mRNA expression of γ-IP10. Results are expressed as mean ± SEM (n = 3 or 4). HBD, heparin-binding domain; ICR, Institute of Cancer Research; IFN, interferon.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4362348&req=5

fig4: Dose-dependent IFNγ expression and IFNγ induced gene expression in mouse liver. ICR mice were injected with different doses of pCpG-IFNγ (solid circles), pCpG-IFNγ-HBD1 (open circles), pCpG-IFNγ-HBD2 (open triangles), and pCpG-IFNγ-HBD3 (open squares) by hydrodynamic gene transfer. Three days after gene transfer, mice were sacrificed and the livers were collected. (a) The mRNA expression of IFNγ, (b) The mRNA expression of SOCS1, (c) The mRNA expression of γ-IP10. Results are expressed as mean ± SEM (n = 3 or 4). HBD, heparin-binding domain; ICR, Institute of Cancer Research; IFN, interferon.
Mentions: pCpG vectors were used instead of pcDNA3 vectors for animal studies, because the former are more efficient in terms of transgene expression than the latter in mice. Mice received a hydrodynamic injection of any of the plasmids at different doses, and the livers were harvested 3 days after injection. Figure 4 shows the mRNA expression of IFNγ (or IFNγ-HBD fusion proteins), a suppressor of cytokine signaling 1 (SOCS1), and IFNγ inducible protein 10 (γ-IP10) in the mouse liver. The mRNA expression of the transgenes was dose dependent in all the groups and did not differ markedly from one to the other. The expression of IFNγ significantly increased the mRNA expression of SOCS1 and γ-IP10, indicating that IFNγ expressed in the liver is biologically active. The expression of these genes in the pCpG-IFNγ-HBD1–injected group was increased to a similar level in the pCpG-IFNγ–injected group. On the other hand, the expression in the pCpG-IFNγ-HBD2– and pCpG-IFNγ-HBD3–injected groups was significantly lower than that in the pCpG-IFNγ– or pCpG-IFNγ-HBD1–injected group, and the mRNA expression in the pCpG-IFNγ-HBD3–injected group was almost negligible over the dose range examined.

Bottom Line: The serum concentration of IFNγ-HBD2 and IFNγ-HBD3 after gene delivery was lower than that of IFNγ or IFNγ-HBD1.Gene delivery of IFNγ-HBD2, but not of IFNγ-HBD3, effectively increased the mRNA expression of IFNγ-inducible genes in the liver, suggesting liver-specific distribution of IFNγ-HBD2.Gene delivery of IFNγ-HBD2-suppressed tumor growth in the liver as efficiently as that of IFNγ with much less symptoms of adverse effects.

View Article: PubMed Central - PubMed

Affiliation: Department of Biopharmaceutics and Drug Metabolism , Graduate School of Pharmaceutical Sciences, Kyoto University , Sakyo-ku, Kyoto, Japan.

ABSTRACT
Sustained gene delivery of interferon (IFN) γ can be an effective treatment, but our previous study showed high levels of IFNγ-induced adverse events, including the loss of body weight. These unwanted events could be reduced by target-specific delivery of IFNγ after in vivo gene transfer. To achieve this, we selected the heparin-binding domain (HBD) of extracellular superoxide dismutase as a molecule to anchor IFNγ to the cell surface. We designed three IFNγ derivatives, IFNγ-HBD1, IFNγ-HBD2, and IFNγ-HBD3, each of which had 1, 2, or 3 HBDs, respectively. Each plasmid-encoding fusion proteins was delivered to the liver, a model target in this study, by hydrodynamic tail vein injection. The serum concentration of IFNγ-HBD2 and IFNγ-HBD3 after gene delivery was lower than that of IFNγ or IFNγ-HBD1. Gene delivery of IFNγ-HBD2, but not of IFNγ-HBD3, effectively increased the mRNA expression of IFNγ-inducible genes in the liver, suggesting liver-specific distribution of IFNγ-HBD2. Gene delivery of IFNγ-HBD2-suppressed tumor growth in the liver as efficiently as that of IFNγ with much less symptoms of adverse effects. These results indicate that the adverse events of IFNγ gene transfer can be prevented by gene delivery of IFNγ-HBD2, a fusion protein with high cell surface affinity.

No MeSH data available.


Related in: MedlinePlus