Growth Retardation of Poorly Transfectable Tumor by Multiple Injections of Plasmids Encoding PE40 Based Targeted Toxin Complexed with Polyethylenimine

Author(s): Yuriy Khodarovich*, Darya Rakhmaninova, German Kagarlitskiy, Anastasia Baryshnikova, Sergey Deyev

Journal Name: Current Gene Therapy

Volume 20 , Issue 4 , 2020

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Graphical Abstract:


Background: One of the approaches to cancer gene therapy relies on tumor transfection with DNA encoding toxins under the control of tumor-specific promoters.

Methods: Here, we used DNA plasmids encoding very potent anti-ERBB2 targeted toxin, driven by the human telomerase promoter or by the ubiquitous CAG promoter (pTERT-ETA and pCAG-ETA) and linear polyethylenimine to target cancer cells.

Results: We showed that the selectivity of cancer cell killing by the pTERT-ETA plasmid is highly dependent upon the method of preparation of DNA-polyethylenimine complexes. After adjustment of complex preparation protocol, cell lines with high activity of telomerase promoter can be selectively killed by transfection with the pTERT-ETA plasmid. We also showed that cells transfected with pTERT-ETA and pCAG-ETA plasmids do not exert any detectable bystander effect in vitro.

Conclusion: Despite this, three intratumoral injections of a plasmid-polyethylenimine complex resulted in substantial growth retardation of a poorly transfectable D2F2/E2 tumor in mice. There were no significant differences in anti-tumor properties between DNA constructs with telomerase or CAG promoters in vivo.

Keywords: Gene therapy, pseudomonas exotoxin A, PE40, targeted therapy, transfection, polyethylenimine.

Lin B, Gao A, Zhang R, et al. Use of a novel integrase-deficient lentivirus for targeted anti-cancer therapy with survivin promoter-driven diphtheria toxin A. Medicine (Baltimore) 2015; 94(31)e1301
Candolfi M, Xiong W, Yagiz K, et al. Gene therapy-mediated delivery of targeted cytotoxins for glioma therapeutics. Proc Natl Acad Sci USA 2010; 107(46): 20021-6.
Liu X, Li J, Tian Y, et al. Enhanced pancreatic cancer gene therapy by combination of adenoviral vector expressing c-erb-B2 (Her-2/neu)-targeted immunotoxin with a replication-competent adenovirus or etoposide. Hum Gene Ther 2010; 21(2): 157-70.
Cemazar M, Sersa G, Wilson J, et al. Effective gene transfer to solid tumors using different nonviral gene delivery techniques: Electroporation, liposomes, and integrin-targeted vector. Cancer Gene Ther 2002; 9: 399-406.
Sokolova E, Proshkina G, Kutova O, et al. Recombinant targeted toxin based on HER2-specific DARPin possesses a strong selective cytotoxic effect in vitro and a potent antitumor activity in vivo. J Control Release 2016; 233: 48-56.
Yamaizumi M, Mekada E, Uchida T, Okada Y. One molecule of diphtheria toxin fragment A introduced into a cell can kill the cell. Cell 1978; 15: 245-50.
Yerushalmi N, Brinkmann U, Brinkmann E, Pai L, Pastan I. Attenuating the growth of tumors by intratumoral administration of DNA encoding Pseudomonas exotoxin via cationic liposomes. Cancer Gene Ther 2000; 7(1): 91-6.
Kimchi-Sarfaty C, Vieira WD, Dodds D, et al. SV40 Pseudovirion gene delivery of a toxin to treat human adenocarcinomas in mice. Cancer Gene Ther 2006; 13(7): 648-57.
Showalter SL, Huang YH, Witkiewicz A, et al. Nanoparticulate delivery of diphtheria toxin DNA effectively kills Mesothelin expressing pancreatic cancer cells. Cancer Biol Ther 2008; 7(10): 1584-90.
Tholey RM, Lal S, Jimbo M, et al. MUC1 promoter-driven DTA as a targeted therapeutic strategy against pancreatic cancer. Mol Cancer Res 2015; 13(3): 439-48.
Michalska M, Wolf P. Pseudomonas exotoxin A: optimized by evolution for effective killing. Front Microbiol 2015; 6: 963.
Stuckey DW, Hingtgen SD, Karakas N, Rich BE, Shah K. Engineering toxin-resistant therapeutic stem cells to treat brain tumors. Stem Cells 2015; 33(2): 589-600.
Kim NW, Piatyszek MA, Prowse KR, et al. Specific association of human telomerase activity with immortal cells and cancer. Science 1994; 266(5193): 2011-5.
Elias A, Gritsenko N, Gorovits R, et al. Anti-cancer binary system activated by bacteriophage HK022 integrase. Oncotarget 2018; 9(44): 27487-501.
Nissim L, Bar-Ziv RH. A tunable dual-promoter integrator for targeting of cancer cells. Mol Syst Biol 2010; 6: 444.
Khodarovich YM, Rakhmaninova DD, Barishnikova AM, Deyev SM. Doxycycline sensitive two-promoter integrator based on the TET-ON 3G Transactivator. Mol Biol (Mosk) 2020; 54(2): 308-12.
Amit D, Hochberg A. Development of targeted therapy for bladder cancer mediated by a double promoter plasmid expressing diphtheria toxin under the control of H19 and IGF2-P4 regulatory sequences. J Transl Med 2010; 8: 134.
Rock KL, Lai JJ, Kono H. Innate and adaptive immune responses to cell death. Immunol Rev 2011; 243(1): 191-205.

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Article Details

Year: 2020
Page: [289 - 296]
Pages: 8
DOI: 10.2174/1566523220999200817101422
Price: $65

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