Abstract
Recent research indicates that cell-mediated gene therapy can be an interesting method to obtain intratumoral expression of therapeutic proteins. This paper explores the possibility of using transfected myeloid-derived suppressor cells (MDSCs), derived from a murine cell line, as cellular vehicles for transporting plasmid DNA (pDNA) encoding interleukin-12 (IL-12) to tumors. Transfecting these cells via electroporation caused massive cell death. This was not due to electroporation-induced cell damage, but was mainly the result of the intracellular presence of plasmids. In contrast, pDNA transfection using Lipofectamine 2000 (LF2000) did not result in a significant loss of viability. Differences in delivery mechanism may explain the distinctive effects on cell viability. Indeed, electroporation is expected to cause a rapid and massive influx of pDNA resulting in cytosolic pDNA levels that most likely surpass the activation threshold of the intracellular DNA sensors leading to cell death. In contrast, a more sustained intracellular release of the pDNA is expected with LF2000. After lipofection with LF2000, 56% of the MDSCs were transfected and transgene expression lasted for at least 24 h. Moreover, biologically relevant amounts of IL-12 were produced by the MDSCs after lipofection with an IL-12 encoding pDNA. In addition, IL-12 transfection caused a significant upregulation of CD80 and considerably reduced the immunosuppressive capacity of the MDSCs. IL-12-transfected MDSCs were still able to migrate to tumor cells, albeit that lipofection of the MDSCs seemed to slightly decrease their migration capacity.
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References
Talmadge JE, Gabrilovich DI . History of myeloid-derived suppressor cells. Nat Rev Cancer 2013; 13: 739–752.
Eisenstein S, Coakley Ba, Briley-Saebo K, Ma G, Chen HM, Meseck M et al. Myeloid-derived suppressor cells as a vehicle for tumor-specific oncolytic viral therapy. Cancer Res 2013; 73: 5003–5015.
Basel MT, Shrestha TB, Bossmann SH, Troyer DL . Cells as delivery vehicles for cancer therapeutics. Ther Deliv 2014; 5: 555–567.
Podhajcer OL, Lopez MV, Mazzolini G . Cytokine gene transfer for cancer therapy. Cytokine Growth Factor Rev 2007; 18: 183–194.
Scanlon KJ . Cancer gene therapy: challenges and opportunities. Anticancer Res 2004; 24: 501–504.
Hu YL, Fu YH, Tabata Y, Gao JQ . Mesenchymal stem cells: a promising targeted-delivery vehicle in cancer gene therapy. J Control Release 2010; 147: 154–162.
Djouad F, Bony C, Apparailly F, Louis-Plence P, Jorgensen C, Noel D . Earlier onset of syngeneic tumors in the presence of mesenchymal stem cells. Transplantation 2006; 82: 1060–1066.
Rubio D, Garcia-Castro J, Martin MC, de la Fuente R, Cigudosa JC, Lloyd AC et al. Spontaneous human adult stem cell transformation. Cancer Res 2005; 65: 3035–3039.
Centeno CJ, Al-Sayegh H, Freeman MD, Smith J, Murrell WD, Bubnov R . A multi-center analysis of adverse events among two thousand, three hundred and seventy two adult patients undergoing adult autologous stem cell therapy for orthopaedic conditions. Int Orthop 2016; 40: 1755–1765.
Ankrum J, Karp JM . Mesenchymal stem cell therapy: two steps forward, one step back. Trends Mol Med 2010; 16: 203–209.
De Becker A, Riet IV . Homing and migration of mesenchymal stromal cells: how to improve the efficacy of cell therapy? World J Stem Cells 2016; 8: 73–87.
Devine SM, Cobbs C, Jennings M, Bartholomew A, Hoffman R . Mesenchymal stem cells distribute to a wide range of tissues following systemic infusion into nonhuman primates. Blood 2003; 101: 2999–3001.
Mosna F, Sensebe L, Krampera M . Human bone marrow and adipose tissue mesenchymal stem cells: a user's guide. Stem Cells Dev 2010; 19: 1449–1470.
Pan PY, Chen HM, Chen SH . Myeloid-derived suppressor cells as a Trojan horse: a cellular vehicle for the delivery of oncolytic viruses. Oncoimmunology 2013; 2: e25083.
Chandra D, Jahangir A, Quispe-Tintaya W, Einstein MH, Gravekamp C . Myeloid-derived suppressor cells have a central role in attenuated Listeria monocytogenes-based immunotherapy against metastatic breast cancer in young and old mice. Br J Cancer 2013; 108: 2281–2290.
Steding CE, Wu ST, Zhang Y, Jeng MH, Elzey BD, Kao C . The role of interleukin-12 on modulating myeloid-derived suppressor cells, increasing overall survival and reducing metastasis. Immunology 2011; 133: 221–238.
Thaci B, Ahmed AU, Ulasov IV, Wainwright DA, Nigam P, Auffinger B et al. Depletion of myeloid-derived suppressor cells during interleukin-12 immunogene therapy does not confer a survival advantage in experimental malignant glioma. Cancer Gene Ther 2014; 21: 38–44.
Del Vecchio M, Bajetta E, Canova S, Lotze MT, Wesa A, Parmiani G et al. Interleukin-12: biological properties and clinical application. Clin Cancer Res 2007; 13: 4677–4685.
Cemazar M, Jarm T, Sersa G . Cancer electrogene therapy with interleukin-12. Curr Gene Ther 2010; 10: 300–311.
Liechtenstein T, Perez-Janices N, Gato M, Caliendo F, Kochan G, Blanco-Luquin I et al. A highly efficient tumor-infiltrating MDSC differentiation system for discovery of anti-neoplastic targets, which circumvents the need for tumor establishment in mice. Oncotarget 2014; 5: 7843–7857.
Apolloni E, Bronte V, Mazzoni A, Serafini P, Cabrelle A, Segal DM et al. Immortalized myeloid suppressor cells trigger apoptosis in antigen-activated T lymphocytes. J Immunol 2000; 165: 6723–6730.
Denies S, Cicchelero L, Polis I, Sanders NN . Immunogenicity and safety of xenogeneic vascular endothelial growth factor receptor-2 DNA vaccination in mice and dogs. Oncotarget 2016; 7: 10905–10916.
Shimokawa T, Okumura K, Ra C . DNA induces apoptosis in electroporated human promonocytic cell line U937. Biochem Biophys Res Commun 2000; 270: 94–99.
Floros T, Tarhini AA . Anticancer cytokines: biology and clinical effects of interferon-alpha2, interleukin (IL)-2, IL-15, IL-21, and IL-12. Semin Oncol 2015; 42: 539–548.
Xu Y, Zhao W, Wu D, Xu J, Lin S, Tang K et al. Isolation of myeloid-derived suppressor cells subsets from spleens of orthotopic liver cancer-bearing mice by fluorescent-activated and magnetic-activated cell sorting: similarities and differences. Int J Clin Exp Pathol 2014; 7: 7545–7553.
Heller L, Todorovic V, Cemazar M . Electrotransfer of single-stranded or double-stranded DNA induces complete regression of palpable B16.F10 mouse melanomas. Cancer Gene Ther 2013; 20: 695–700.
Stacey KJ, Ross IL, Hume DA . Electroporation and DNA-dependent cell death in murine macrophages. Immunol Cell Biol 1993; 71 (Pt 2): 75–85.
Hemmi H, Takeuchi O, Kawai T, Kaisho T, Sato S, Sanjo H et al. A Toll-like receptor recognizes bacterial DNA. Nature 2000; 408: 740–745.
Znidar K, Bosnjak M, Cemazar M, Heller LC, Cytosolic DNA . Sensor upregulation accompanies DNA electrotransfer in B16.F10 melanoma cells. Mol Ther Nucleic Acids 2016; 5: e322.
Magnusson T, Haase R, Schleef M, Wagner E, Ogris M . Sustained, high transgene expression in liver with plasmid vectors using optimized promoter-enhancer combinations. J Gene Med 2011; 13: 382–391.
Brooks AR, Harkins RN, Wang P, Qian HS, Liu P, Rubanyi GM . Transcriptional silencing is associated with extensive methylation of the CMV promoter following adenoviral gene delivery to muscle. J Gene Med 2004; 6: 395–404.
Tolmachov OE, Subkhankulova T, Tolmachova T . Silencing of transgene expression: a gene therapy perspective. In: Martin-Molina F (ed.) Gene Therapy - Tools and Potential Applications. InTech: Rijeka, Croatia, 2013, pp 49–68.
Wei LZ, Xu Y, Nelles EM, Furlonger C, Wang JC, Di Grappa MA et al. Localized interleukin-12 delivery for immunotherapy of solid tumours. J Cell Mol Med 2013; 17: 1465–1474.
Zhang L, Morgan Ra, Beane JD, Zheng Z, Dudley ME, Kassim SH et al. Tumor infiltrating lymphocytes genetically engineered with an inducible gene encoding interleukin-12 for the immunotherapy of metastatic melanoma. Clin Cancer Res 2015; 21: 2278–2288.
Kerkar SP, Goldszmid RS, Muranski P, Chinnasamy D, Yu Z, Reger RN et al. IL-12 triggers a programmatic change in dysfunctional myeloid-derived cells within mouse tumors. J Clin Invest 2011; 121: 4746–4757.
Manzotti CN, Liu MK, Burke F, Dussably L, Zheng Y, Sansom DM . Integration of CD28 and CTLA-4 function results in differential responses of T cells to CD80 and CD86. Eur J Immunol 2006; 36: 1413–1422.
Denies S, Cicchelero L, Van Audenhove I, Sanders NN . Combination of interleukin-12 gene therapy, metronomic cyclophosphamide and DNA cancer vaccination directs all arms of the immune system towards tumor eradication. J Control Release 2014; 187: 175–182.
Ichikawa M, Williams R, Wang L, Vogl T, Srikrishna G . S100A8/A9 activate key genes and pathways in colon tumor progression. Mol Cancer Res 2011; 9: 133–148.
Hermani A, De Servi B, Medunjanin S, Tessier PA, Mayer D . S100A8 and S100A9 activate MAP kinase and NF-kappaB signaling pathways and trigger translocation of RAGE in human prostate cancer cells. Exp Cell Res 2006; 312: 184–197.
Zheng R, Chen S, Chen S . Correlation between myeloid-derived suppressor cells and S100A8/A9 in tumor and autoimmune diseases. Int Immunopharmacol 2015; 29: 919–925.
Garcia-Hernandez ML, Hernandez-Pando R, Gariglio P, Berumen J . Interleukin-10 promotes B16-melanoma growth by inhibition of macrophage functions and induction of tumour and vascular cell proliferation. Immunology 2002; 105: 231–243.
Uekusa Y, Gao P, Yamaguchi N, Tomura M, Mukai T, Nakajima C et al. A role for endogenous IL-12 in tumor immunity: IL-12 is required for the acquisition of tumor-migratory capacity by T cells and the development of T cell-accepting capacity in tumor masses. J Leukoc Biol 2002; 72: 864–873.
Rabinowich H, Herberman RB, Whiteside TL . Differential effects of IL12 and IL2 on expression and function of cellular adhesion molecules on purified human natural killer cells. Cell Immunol 1993; 152: 481–498.
Acknowledgements
This work was supported by grants of the University of Ghent (BOF) and by grants from the research foundation—Flanders (FWO; G.0235.11N and G.0621.10N). FC is a fellow of the research foundation—Flanders (FWO).
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Denies, S., Combes, F., Ghekiere, C. et al. In vitro exploration of a myeloid-derived suppressor cell line as vehicle for cancer gene therapy. Cancer Gene Ther 24, 149–155 (2017). https://doi.org/10.1038/cgt.2016.60
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DOI: https://doi.org/10.1038/cgt.2016.60
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