Abstract
Sustained expression of murine interferon (IFN)-γ (Muγ) was found to be effective in preventing tumor metastasis and atopic dermatitis in mouse models. However, our preliminary experiments suggested that the time-dependent decrease in the Muγ expression was not compensated for by repeated injections of Muγ-expressing plasmid. To identify the mechanism underlying this observation, a reporter plasmid was hydrodynamically injected into mice and the levels of the plasmid, mRNA and reporter protein were measured in mice receiving a pre- or co-administration of Muγ-expressing plasmid. Co-injection of Muγ-expressing plasmid had no significant effects on transgene expression from the reporter plasmid. In contrast, pre-injection of Muγ-expressing plasmid greatly inhibited the expression of the reporter protein. Moreover, pre-injection of Muγ-expressing plasmid also reduced the amount of the reporter plasmid in the nuclear fraction of mouse liver to <10%, and that of reporter mRNA to <1%. The degree of reduction in the expression of reporter protein was comparable with the reduction in mRNA. These results indicate that the difficulty in regaining the expression level of IFN-γ is due to the impaired delivery of plasmid to the nucleus and to the suppression of transcription from the plasmid.
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References
Goodbourn S, Didcock L, Randall RE . Interferons: cell signalling, immune modulation, antiviral response and virus countermeasures. J Gen Virol 2000; 81: 2341–2364.
Samuel CE . Antiviral actions of interferons. Clin Microbiol Rev 2001; 14: 778–809.
Grassegger A, Hopfl R . Significance of the cytokine interferon gamma in clinical dermatology. Clin Exp Dermatol 2004; 29: 584–588.
Smyth MJ, Cretney E, Kershaw MH, Hayakawa Y . Cytokines in cancer immunity and immunotherapy. Immunol Rev 2004; 202: 275–293.
Borden EC, Sen GC, Uze G, Silverman RH, Ransohoff RM, Foster GR et al. Interferons at age 50: past, current and future impact on biomedicine. Nat Rev Drug Discov 2007; 6: 975–990.
Pepinsky RB, LePage DJ, Gill A, Chakraborty A, Vaidyanathan S, Green M et al. Improved pharmacokinetic properties of a polyethylene glycol-modified form of interferon-beta-1a with preserved in vitro bioactivity. J Pharmacol Exp Ther 2001; 297: 1059–1066.
Palumbo E . PEG-interferon alpha-2b for acute hepatitis C: a review. Mini Rev Med Chem 2007; 7: 839–843.
Mata-Espinosa DA, Mendoza-Rodríguez V, Aguilar-León D, Rosales R, López-Casillas F, Hernández-Pando R . Therapeutic effect of recombinant adenovirus encoding interferon-gamma in a murine model of progressive pulmonary tuberculosis. Mol Ther 2008; 16: 1065–1072.
Shiau AL, Lin CY, Tzai TS, Wu CL . Postoperative immuno-gene therapy of murine bladder tumor by in vivo administration of retroviruses expressing mouse interferon-gamma. Cancer Gene Ther 2001; 8: 73–81.
Kobayashi N, Kuramoto T, Chen S, Watanabe Y, Takakura Y . Therapeutic effect of intravenous interferon gene delivery with naked plasmid DNA in murine metastasis models. Mol Ther 2002; 6: 737–744.
Kawano H, Nishikawa M, Mitsui M, Takahashi Y, Kako K, Yamaoka K et al. Improved anti-cancer effect of interferon gene transfer by sustained expression using CpG-reduced plasmid DNA. Int J Cancer 2007; 121: 401–406.
Mitsui M, Nishikawa M, Zang L, Ando M, Hattori K, Takahashi Y et al. Effect of the content of unmethylated CpG dinucleotides in plasmid DNA on the sustainability of transgene expression. J Gene Med 2009; 11: 435–443.
Liu F, Song Y, Liu D . Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA. Gene Therapy 1999; 6: 1258–1266.
Hattori K, Nishikawa M, Watcharanurak K, Ikoma A, Kabashima K, Toyota H et al. Sustained exogenous expression of therapeutic levels of IFN-gamma ameliorates atopic dermatitis in NC/Nga mice via Th1 polarization. J Immunol 2010; 184: 2729–2735.
Nishikawa M, Nakayama A, Takahashi Y, Fukuhara Y, Takakura Y . Reactivation of silenced transgene expression in mouse liver by rapid, large-volume injection of isotonic solution. Hum Gene Ther 2008; 19: 1009–1020.
Qin L, Ding Y, Pahud DR, Chang E, Imperiale MJ, Bromberg JS . Promoter attenuation in gene therapy: interferon-gamma and tumor necrosis factor-alpha inhibit transgene expression. Hum Gene Ther 1997; 8: 2019–2029.
Harms JS, Splitter GA . Interferon-gamma inhibits transgene expression driven by SV40 or CMV promoters but augments expression driven by the mammalian MHC I promoter. Hum Gene Ther 1995; 6: 1291–1297.
Ghazizadeh S, Carroll JM, Taichman LB . Repression of retrovirus-mediated transgene expression by interferons: implications for gene therapy. J Virol 1997; 71: 9163–9169.
Acsadi G, O'Hagan D, Lochmüller H, Prescott S, Larochelle N, Nalbantoglu J et al. Interferons impair early transgene expression by adenovirus-mediated gene transfer in muscle cells. J Mol Med 1998; 76: 442–450.
Lui VW, Falo Jr LD, Huang L . Systemic production of IL-12 by naked DNA mediated gene transfer: toxicity and attenuation of transgene expression in vivo. J Gene Med 2001; 3: 384–393.
Sung RS, Qin L, Bromberg JS . TNFalpha and IFNgamma induced by innate anti-adenoviral immune responses inhibit adenovirus-mediated transgene expression. Mol Ther 2001; 3: 757–767.
Luo P, Reed BD, Tsang TC, Harris DT, Flavell RA . A self-augmenting gene expression cassette for enhanced and sustained transgene expression in the presence of proinflammatory cytokines. DNA Cell Biol 2006; 25: 659–667.
Reboredo M, Zabala M, Mauleon I, DeLas Rivas J, Kreppel F, Kochanek S et al. Interleukin-12 inhibits liver-specific drug-inducible systems in vivo. Gene Therapy 2008; 15: 277–288.
Song YK, Liu F, Chu S, Liu D . Characterization of cationic liposome-mediated gene transfer in vivo by intravenous administration. Hum Gene Ther 1997; 8: 1585–1594.
Li S, Wu SP, Whitmore M, Loeffert EJ, Wang L, Watkins SC et al. Effect of immune response on gene transfer to the lung via systemic administration of cationic lipidic vectors. Am J Physiol 1999; 276: 796–804.
Yasuda S, Yoshida H, Nishikawa M, Takakura Y . Comparison of the type of liposome involving cytokine production induced by non-CpG Lipoplex in macrophages. Mol Pharm 2010; 7: 533–542.
Yoshida H, Nishikawa M, Yasuda S, Mizuno Y, Toyota H, Kiyota T et al. TLR9-dependent systemic interferon-beta production by intravenous injection of plasmid DNA/cationic liposome complex in mice. J Gene Med 2009; 11: 708–717.
Yew NS, Zhao H, Wu IH, Song A, Tousignant JD, Przybylska M et al. Reduced inflammatory response to plasmid DNA vectors by elimination and inhibition of immunostimulatory CpG motifs. Mol Ther 2000; 1: 255–262.
Smolarczyk R, Cichoń T, Sochanik A, Szala S . Negligible induction of IFN-gamma, IL-12 and TNF-alpha by DNA-PEI 750 kDa/albumin complexes. Cytokine 2005; 29: 283–287.
Ramana CV, Grammatikakis N, Chernov M, Nguyen H, Goh KC, Williams BR et al. Regulation of c-myc expression by IFN-gamma through Stat1-dependent and -independent pathways. EMBO J 2000; 19: 263–272.
Schaefer A, Unterberger C, Frankenberger M, Lohrum M, Staples KJ, Werner T et al. Mechanism of interferon-gamma mediated down-regulation of interleukin-10 gene expression. Mol Immunol 2009; 46: 1351–1359.
Kobayashi N, Nishikawa M, Hirata K, Takakura Y . Hydrodynamics-based procedure involves transient hyperpermeability in the hepatic cellular membrane: implication of a nonspecific process in efficient intracellular gene delivery. J Gene Med 2004; 6: 584–592.
Zhang G, Gao X, Song YK, Vollmer R, Stolz DB, Gasiorowski JZ et al. Hydroporation as the mechanism of hydrodynamic delivery. Gene Therapy 2004; 11: 675–682.
Dean DA, Dean BS, Muller S, Smith LC . Sequence requirements for plasmid nuclear import. Exp Cell Res 1999; 253: 713–722.
Roeder RG . The role of general initiation factors in transcription by RNA polymerase II. Trends Biochem Sci 1996; 21: 327–335.
Deng WG, Montero AJ, Wu KK . Interferon-γ suppresses cyclooxygenase-2 promoter activity by inhibiting C-Jun and C/EBPβ binding. Arterioscler Thromb Vasc Biol 2007; 27: 1752–1759.
Ganster RW, Guo Z, Shao L, Geller DA . Differential effects of TNF-alpha and IFN-gamma on gene transcription mediated by NF-kappaB-Stat1 interactions. J Interferon Cytokine Res 2005; 25: 707–719.
Shao L, Guo Z, Geller DA . Transcriptional suppression of cytokine-induced iNOS gene expression by IL-13 through IRF-1/ISRE signaling. Biochem Biophys Res Commun 2007; 362: 582–586.
Nomura T, Yasuda K, Yamada T, Okamoto S, Mahato RI, Watanabe Y et al. Gene expression and antitumor effects following direct interferon (IFN)-gamma gene transfer with naked plasmid DNA and DC-chol liposome complexes in mice. Gene Therapy 1999; 6: 121–129.
Mir LM, Bureau MF, Gehl J, Rangara R, Rouy D, Caillaud JM et al. High-efficiency gene transfer into skeletal muscle mediated by electric pulses. Proc Natl Acad Sci USA 1999; 96: 4262–4267.
Ochiai H, Harashima H, Kamiya H . Intranuclear disposition of exogenous DNA in vivo: silencing, methylation and fragmentation. FEBS Lett 2006; 580: 918–922.
Ochiai H, Fujimuro M, Yokosawa H, Harashima H, Kamiya H . Transient activation of transgene expression by hydrodynamics-based injection may cause rapid decrease in plasmid DNA expression. Gene Therapy 2007; 14: 1152–1159.
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This work was supported in part by Grants-in-Aid for Scientific Research (B) and for Challenging Exploratory Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan.
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Zang, L., Nishikawa, M., Machida, K. et al. Inhibition of nuclear delivery of plasmid DNA and transcription by interferon γ: hurdles to be overcome for sustained gene therapy. Gene Ther 18, 891–897 (2011). https://doi.org/10.1038/gt.2011.35
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DOI: https://doi.org/10.1038/gt.2011.35
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