1887

Abstract

In patients with malignancy, the major barrier to achieving complete response is emergence of resistance to current chemotherapeutic agents. One of the major mechanisms by which tumour cells become resistant to therapies is by altering cellular drug targets through mutations and/or deletions. Resistance by this mechanism is achieved more easily if the drug has limited cellular targets and/or processes. We hypothesized that as exotoxin T (ExoT) targets six proteins that are required for cancer cell survival and proliferation, it is highly unlikely for cancer cells to develop resistance to this toxin. We assessed ExoT’s cytotoxicity against multiple invasive and highly resistant tumour cell lines in order to evaluate its potential as a chemotherapeutic agent. Our data demonstrated that ExoT induced potent cytotoxicity in all tumour cell lines that we examined. Collectively, our data highlighted the potential of ExoT as a possible chemotherapeutic candidate for the treatment of cancer.

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2015-02-01
2020-01-28
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References

  1. Alcazar O., Achberger S., Aldrich W., Hu Z., Negrotto S., Saunthararajah Y., Triozzi P.. ( 2011;). Epigenetic regulation by decitabine of melanoma differentiation in vitro and in vivo. . Int J Cancer 131:, 18–29. [CrossRef][PubMed]
    [Google Scholar]
  2. Arechabala B., Coiffard C., Rivalland P., Coiffard L. J. M., de Roeck-Holtzhauer Y.. ( 1999;). Comparison of cytotoxicity of various surfactants tested on normal human fibroblast cultures using the neutral red test, MTT assay and LDH release. . J Appl Toxicol 19:, 163–165. [CrossRef][PubMed]
    [Google Scholar]
  3. Aslakson C. J., Miller F. R.. ( 1992;). Selective events in the metastatic process defined by analysis of the sequential dissemination of subpopulations of a mouse mammary tumor. . Cancer Res 52:, 1399–1405.[PubMed]
    [Google Scholar]
  4. Atkins M. B., Hsu J., Lee S., Cohen G. I., Flaherty L. E., Sosman J. A., Sondak V. K., Kirkwood J. M..Eastern Cooperative Oncology Group ( 2008;). Phase III trial comparing concurrent biochemotherapy with cisplatin, vinblastine, dacarbazine, interleukin-2, and interferon alfa-2b with cisplatin, vinblastine, and dacarbazine alone in patients with metastatic malignant melanoma (E3695): a trial coordinated by the Eastern Cooperative Oncology Group. . J Clin Oncol 26:, 5748–5754. [CrossRef][PubMed]
    [Google Scholar]
  5. Bagrodia S., Taylor S. J., Jordon K. A., Van Aelst L., Cerione R. A.. ( 1998;). A novel regulator of p21-activated kinases. . J Biol Chem 273:, 23633–23636. [CrossRef][PubMed]
    [Google Scholar]
  6. Baguley B. C.. ( 2010;). Multiple drug resistance mechanisms in cancer. . Mol Biotechnol 46:, 308–316. [CrossRef][PubMed]
    [Google Scholar]
  7. Becker N., Benhar I.. ( 2012;). Antibody-based immunotoxins for the treatment of cancer. . Antibodies 1:, 39–69. [CrossRef]
    [Google Scholar]
  8. Bertram J. S., Janik P.. ( 1980;). Establishment of a cloned line of Lewis lung carcinoma cells adapted to cell culture. . Cancer Lett 11:, 63–73. [CrossRef][PubMed]
    [Google Scholar]
  9. Breitbach C. J., Burke J., Jonker D., Stephenson J., Haas A. R., Chow L. Q., Nieva J., Hwang T. H., Moon A. et al. ( 2011;). Intravenous delivery of a multi-mechanistic cancer-targeted oncolytic poxvirus in humans. . Nature 477:, 99–102. [CrossRef][PubMed]
    [Google Scholar]
  10. Cailleau R., Young R., Olivé M., Reeves W. J. Jr. ( 1974;). Breast tumor cell lines from pleural effusions. . J Natl Cancer Inst 53:, 661–674.[PubMed]
    [Google Scholar]
  11. Calcagno A. M., Ambudkar S. V.. ( 2010;). Molecular mechanisms of drug resistance in single-step and multi-step drug-selected cancer cells. . Methods Mol Biol 596:, 77–93. [CrossRef][PubMed]
    [Google Scholar]
  12. Chaudhary V. K., FitzGerald D. J., Adhya S., Pastan I.. ( 1987;). Activity of a recombinant fusion protein between transforming growth factor type alpha and Pseudomonas toxin. . Proc Natl Acad Sci U S A 84:, 4538–4542. [CrossRef][PubMed]
    [Google Scholar]
  13. Chaudhary V. K., Queen C., Junghans R. P., Waldmann T. A., FitzGerald D. J., Pastan I.. ( 1989;). A recombinant immunotoxin consisting of two antibody variable domains fused to Pseudomonas exotoxin. . Nature 339:, 394–397. [CrossRef][PubMed]
    [Google Scholar]
  14. Chen W. J., Xiong Z. A., Tang Y., Dong P. T., Li P., Wang Z. G.. ( 2012;). Feasibility and effect of ultrasound microbubble-mediated wild-type p53 gene transfection of HeLa cells. . Exp Ther Med 3:, 999–1004.[PubMed]
    [Google Scholar]
  15. Cho S. Y., Klemke R. L.. ( 2000;). Extracellular-regulated kinase activation and CAS/Crk coupling regulate cell migration and suppress apoptosis during invasion of the extracellular matrix. . J Cell Biol 149:, 223–236. [CrossRef][PubMed]
    [Google Scholar]
  16. Dang L. H., Bettegowda C., Huso D. L., Kinzler K. W., Vogelstein B.. ( 2001;). Combination bacteriolytic therapy for the treatment of experimental tumors. . Proc Natl Acad Sci U S A 98:, 15155–15160. [CrossRef][PubMed]
    [Google Scholar]
  17. Decker T., Lohmann-Matthes M. L.. ( 1988;). A quick and simple method for the quantitation of lactate dehydrogenase release in measurements of cellular cytotoxicity and tumor necrosis factor (TNF) activity. . J Immunol Methods 115:, 61–69. [CrossRef][PubMed]
    [Google Scholar]
  18. del Peso L., Hernández-Alcoceba R., Embade N., Carnero A., Esteve P., Paje C., Lacal J. C.. ( 1997;). Rho proteins induce metastatic properties in vivo. . Oncogene 15:, 3047–3057. [CrossRef][PubMed]
    [Google Scholar]
  19. Feng Y. X., Zhao J. S., Li J. J., Wang T., Cheng S. Q., Yuan Y., Wang F., Wang X. F., Xie D.. ( 2010;). Liver cancer: ephrinA2 promotes tumorigenicity through Rac1/Akt/NF-kappaB signaling pathway. . Hepatology 51:, 535–544. [CrossRef][PubMed]
    [Google Scholar]
  20. Fiers W., Beyaert R., Declercq W., Vandenabeele P.. ( 1999;). More than one way to die: apoptosis, necrosis and reactive oxygen damage. . Oncogene 18:, 7719–7730. [CrossRef][PubMed]
    [Google Scholar]
  21. Flaherty K. T., Infante J. R., Daud A., Gonzalez R., Kefford R. F., Sosman J., Hamid O., Schuchter L., Cebon J. et al. ( 2012;). Combined BRAF and MEK inhibition in melanoma with BRAF V600 mutations. . N Engl J Med 367:, 1694–1703. [CrossRef][PubMed]
    [Google Scholar]
  22. Fogh J., Fogh J. M., Orfeo T.. ( 1977;). One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. . J Natl Cancer Inst 59:, 221–226.[PubMed]
    [Google Scholar]
  23. Foley B. T., Moehring J. M., Moehring T. J.. ( 1995;). Mutations in the elongation factor 2 gene which confer resistance to diphtheria toxin and Pseudomonas exotoxin A. Genetic and biochemical analyses. . J Biol Chem 270:, 23218–23225. [CrossRef][PubMed]
    [Google Scholar]
  24. García Z., Silio V., Marqués M., Cortés I., Kumar A., Hernandez C., Checa A. I., Serrano A., Carrera A. C.. ( 2006;). A PI3K activity-independent function of p85 regulatory subunit in control of mammalian cytokinesis. . EMBO J 25:, 4740–4751. [CrossRef][PubMed]
    [Google Scholar]
  25. Garrity-Ryan L., Shafikhani S., Balachandran P., Nguyen L., Oza J., Jakobsen T., Sargent J., Fang X., Cordwell S. et al. ( 2004;). The ADP ribosyltransferase domain of Pseudomonas aeruginosa ExoT contributes to its biological activities. . Infect Immun 72:, 546–558. [CrossRef][PubMed]
    [Google Scholar]
  26. Gómez J., Martínez C., Giry M., García A., Rebollo A.. ( 1997;). Rho prevents apoptosis through Bcl-2 expression: implications for interleukin-2 receptor signal transduction. . Eur J Immunol 27:, 2793–2799. [CrossRef][PubMed]
    [Google Scholar]
  27. Gonzalez-Angulo A. M., Morales-Vasquez F., Hortobagyi G. N.. ( 2007;). Overview of resistance to systemic therapy in patients with breast cancer. . Adv Exp Med Biol 608:, 1–22. [CrossRef][PubMed]
    [Google Scholar]
  28. Gorre M. E., Mohammed M., Ellwood K., Hsu N., Paquette R., Rao P. N., Sawyers C. L.. ( 2001;). Clinical resistance to STI-571 cancer therapy caused by BCR-ABL gene mutation or amplification. . Science 293:, 876–880. [CrossRef][PubMed]
    [Google Scholar]
  29. Gottesman M. M.. ( 2002;). Mechanisms of cancer drug resistance. . Annu Rev Med 53:, 615–627. [CrossRef][PubMed]
    [Google Scholar]
  30. He X., Liu J., Qi Y., Brakebusch C., Chrostek-Grashoff A., Edgar D., Yurchenco P. D., Corbett S. A., Lowry S. F. et al. ( 2010;). Rac1 is essential for basement membrane-dependent epiblast survival. . Mol Cell Biol 30:, 3569–3581. [CrossRef][PubMed]
    [Google Scholar]
  31. Heasman S. J., Ridley A. J.. ( 2008;). Mammalian Rho GTPases: new insights into their functions from in vivo studies. . Nat Rev Mol Cell Biol 9:, 690–701. [CrossRef][PubMed]
    [Google Scholar]
  32. Homet B., Ribas A.. ( 2013;). New drug targets in metastatic melanoma. . J Pathol 232:, 134–141. [CrossRef][PubMed]
    [Google Scholar]
  33. Hwang T. L., Liang Y., Chien K. Y., Yu J. S.. ( 2006;). Overexpression and elevated serum levels of phosphoglycerate kinase 1 in pancreatic ductal adenocarcinoma. . Proteomics 6:, 2259–2272. [CrossRef][PubMed]
    [Google Scholar]
  34. Jørgensen R., Wang Y., Visschedyk D., Merrill A. R.. ( 2008;). The nature and character of the transition state for the ADP-ribosyltransferase reaction. . EMBO Rep 9:, 802–809. [CrossRef][PubMed]
    [Google Scholar]
  35. Karbach J., Neumann A., Brand K., Wahle C., Siegel E., Maeurer M., Ritter E., Tsuji T., Gnjatic S. et al. ( 2012;). Phase I clinical trial of mixed bacterial vaccine (Coley’s toxins) in patients with NY-ESO-1 expressing cancers: immunological effects and clinical activity. . Clin Cancer Res 18:, 5449–5459. [CrossRef][PubMed]
    [Google Scholar]
  36. Kawakami K., Nakajima O., Morishita R., Nagai R.. ( 2006;). Targeted anticancer immunotoxins and cytotoxic agents with direct killing moieties. . ScientificWorldJournal 6:, 781–790. [CrossRef][PubMed]
    [Google Scholar]
  37. Kazmierczak B. I., Engel J. N.. ( 2002;). Pseudomonas aeruginosa ExoT acts in vivo as a GTPase-activating protein for RhoA, Rac1, and Cdc42. . Infect Immun 70:, 2198–2205. [CrossRef][PubMed]
    [Google Scholar]
  38. Kirn D. H., Thorne S. H.. ( 2009;). Targeted and armed oncolytic poxviruses: a novel multi-mechanistic therapeutic class for cancer. . Nat Rev Cancer 9:, 64–71. [CrossRef][PubMed]
    [Google Scholar]
  39. Kobayashi S., Boggon T. J., Dayaram T., Jänne P. A., Kocher O., Meyerson M., Johnson B. E., Eck M. J., Tenen D. G., Halmos B.. ( 2005;). EGFR mutation and resistance of non-small-cell lung cancer to gefitinib. . N Engl J Med 352:, 786–792. [CrossRef][PubMed]
    [Google Scholar]
  40. Korrer M. J., Routes J. M.. ( 2014;). Possible role of arginase-1 in concomitant tumor immunity. . PLoS One 9:, e91370. [CrossRef][PubMed]
    [Google Scholar]
  41. Korzeniewski C., Callewaert D. M.. ( 1983;). An enzyme-release assay for natural cytotoxicity. . J Immunol Methods 64:, 313–320. [CrossRef][PubMed]
    [Google Scholar]
  42. Krall R., Schmidt G., Aktories K., Barbieri J. T.. ( 2000;). Pseudomonas aeruginosa ExoT is a Rho GTPase-activating protein. . Infect Immun 68:, 6066–6068. [CrossRef][PubMed]
    [Google Scholar]
  43. Kreitman R. J.. ( 2006;). Immunotoxins for targeted cancer therapy. . AAPS J 8:, E532–E551. [CrossRef][PubMed]
    [Google Scholar]
  44. Kreitman R. J.. ( 2009;). Recombinant immunotoxins for the treatment of chemoresistant hematologic malignancies. . Curr Pharm Des 15:, 2652–2664. [CrossRef][PubMed]
    [Google Scholar]
  45. Kroemer G., Galluzzi L., Vandenabeele P., Abrams J., Alnemri E. S., Baehrecke E. H., Blagosklonny M. V., El-Deiry W. S., Golstein P. et al. ( 2009;). Classification of cell death: recommendations of the Nomenclature Committee on Cell Death 2009. . Cell Death Differ 16:, 3–11. [CrossRef][PubMed]
    [Google Scholar]
  46. Lamorte L., Royal I., Naujokas M., Park M.. ( 2002;). Crk adapter proteins promote an epithelial-mesenchymal-like transition and are required for HGF-mediated cell spreading and breakdown of epithelial adherens junctions. . Mol Biol Cell 13:, 1449–1461. [CrossRef][PubMed]
    [Google Scholar]
  47. Lavrik I. N., Golks A., Krammer P. H.. ( 2005;). Caspases: pharmacological manipulation of cell death. . J Clin Invest 115:, 2665–2672. [CrossRef][PubMed]
    [Google Scholar]
  48. Lay A. J., Jiang X. M., Kisker O., Flynn E., Underwood A., Condron R., Hogg P. J.. ( 2000;). Phosphoglycerate kinase acts in tumour angiogenesis as a disulphide reductase. . Nature 408:, 869–873. [CrossRef][PubMed]
    [Google Scholar]
  49. Mahmood F., Hakimiyan A., Jayaraman V., Wood S., Sivaramakrishnan G., Rehman T., Reuhs B. L., Chubinskaya S., Shafikhani S. H.. ( 2013;). A novel human antimicrobial factor targets Pseudomonas aeruginosa through its type III secretion system. . J Med Microbiol 62:, 531–539. [CrossRef][PubMed]
    [Google Scholar]
  50. Murga C., Zohar M., Teramoto H., Gutkind J. S.. ( 2002;). Rac1 and RhoG promote cell survival by the activation of PI3K and Akt, independently of their ability to stimulate JNK and NF-kappaB. . Oncogene 21:, 207–216. [CrossRef][PubMed]
    [Google Scholar]
  51. Overwijk W. W., Restifo N. P.. ( 2001;). B16 as a mouse model for human melanoma. . Curr Protoc Immunol 39:, 20.1.1–20.1.29.[PubMed]
    [Google Scholar]
  52. Park T. J., Boyd K., Curran T.. ( 2006;). Cardiovascular and craniofacial defects in Crk-null mice. . Mol Cell Biol 26:, 6272–6282. [CrossRef][PubMed]
    [Google Scholar]
  53. Park B. H., Hwang T., Liu T. C., Sze D. Y., Kim J. S., Kwon H. C., Oh S. Y., Han S. Y., Yoon J. H. et al. ( 2008;). Use of a targeted oncolytic poxvirus, JX-594, in patients with refractory primary or metastatic liver cancer: a phase I trial. . Lancet Oncol 9:, 533–542. [CrossRef][PubMed]
    [Google Scholar]
  54. Pastan I., Beers R., Bera T. K.. ( 2004;). Recombinant immunotoxins in the treatment of cancer. . Methods Mol Biol 248:, 503–518.[PubMed]
    [Google Scholar]
  55. Patyar S., Joshi R., Byrav D. S., Prakash A., Medhi B., Das B. K.. ( 2010;). Bacteria in cancer therapy: a novel experimental strategy. . J Biomed Sci 17:, 21. [CrossRef][PubMed]
    [Google Scholar]
  56. Perfetto S. P., Chattopadhyay P. K., Lamoreaux L., Nguyen R., Ambrozak D., Koup R. A., Roederer M.. ( 2010;). Amine-reactive dyes for dead cell discrimination in fixed samples. . Curr Protoc Cytom 53:, 9.34.1–9.34.14. [CrossRef][PubMed]
    [Google Scholar]
  57. Reddy A., Caler E. V., Andrews N. W.. ( 2001;). Plasma membrane repair is mediated by Ca2+-regulated exocytosis of lysosomes. . Cell 106:, 157–169. [CrossRef][PubMed]
    [Google Scholar]
  58. Riol-Blanco L., Delgado-Martín C., Sánchez-Sánchez N., Alonso-C L. M., Gutiérrez-López M. D., Del Hoyo G. M., Navarro J., Sánchez-Madrid F., Cabañas C. et al. ( 2009;). Immunological synapse formation inhibits, via NF-kappaB and FOXO1, the apoptosis of dendritic cells. . Nat Immunol 10:, 753–760. [CrossRef][PubMed]
    [Google Scholar]
  59. Rockwell S. C., Kallman R. F., Fajardo L. F.. ( 1972;). Characteristics of a serially transplanted mouse mammary tumor and its tissue-culture-adapted derivative. . J Natl Cancer Inst 49:, 735–749.[PubMed]
    [Google Scholar]
  60. Rodrigues S. P., Fathers K. E., Chan G., Zuo D., Halwani F., Meterissian S., Park M.. ( 2005;). CrkI and CrkII function as key signaling integrators for migration and invasion of cancer cells. . Mol Cancer Res 3:, 183–194.[PubMed]
    [Google Scholar]
  61. Scherer W. F., Syverton J. T., Gey G. O.. ( 1953;). Studies on the propagation in vitro of poliomyelitis viruses. IV. Viral multiplication in a stable strain of human malignant epithelial cells (strain HeLa) derived from an epidermoid carcinoma of the cervix. . J Exp Med 97:, 695–710. [CrossRef][PubMed]
    [Google Scholar]
  62. Shafikhani S. H., Engel J.. ( 2006;). Pseudomonas aeruginosa type III-secreted toxin ExoT inhibits host-cell division by targeting cytokinesis at multiple steps. . Proc Natl Acad Sci U S A 103:, 15605–15610. [CrossRef][PubMed]
    [Google Scholar]
  63. Shafikhani S. H., Morales C., Engel J.. ( 2008;). The Pseudomonas aeruginosa type III secreted toxin ExoT is necessary and sufficient to induce apoptosis in epithelial cells. . Cell Microbiol 10:, 994–1007. [CrossRef][PubMed]
    [Google Scholar]
  64. Sirsi S. R., Borden M. A.. ( 2012;). Advances in ultrasound mediated gene therapy using microbubble contrast agents. . Theranostics 2:, 1208–1222. [CrossRef][PubMed]
    [Google Scholar]
  65. Smith B., Land H.. ( 2012;). Anticancer activity of the cholesterol exporter ABCA1 gene. . Cell Rep 2:, 580–590. [CrossRef][PubMed]
    [Google Scholar]
  66. Smith A. H., Fujii H., Kuliszewski M. A., Leong-Poi H.. ( 2011;). Contrast ultrasound and targeted microbubbles: diagnostic and therapeutic applications for angiogenesis. . J Cardiovasc Transl Res 4:, 404–415. [CrossRef][PubMed]
    [Google Scholar]
  67. Soule H. D., Vazguez J., Long A., Albert S., Brennan M.. ( 1973;). A human cell line from a pleural effusion derived from a breast carcinoma. . J Natl Cancer Inst 51:, 1409–1416.[PubMed]
    [Google Scholar]
  68. Sun J., Barbieri J. T.. ( 2003;). Pseudomonas aeruginosa ExoT ADP-ribosylates CT10 regulator of kinase (Crk) proteins. . J Biol Chem 278:, 32794–32800. [CrossRef][PubMed]
    [Google Scholar]
  69. Van Mellaert L., Barbé S., Anné J.. ( 2006;). Clostridium spores as anti-tumour agents. . Trends Microbiol 14:, 190–196. [CrossRef][PubMed]
    [Google Scholar]
  70. Vanderplasschen A., Mathew E., Hollinshead M., Sim R. B., Smith G. L.. ( 1998;). Extracellular enveloped vaccinia virus is resistant to complement because of incorporation of host complement control proteins into its envelope. . Proc Natl Acad Sci U S A 95:, 7544–7549. [CrossRef][PubMed]
    [Google Scholar]
  71. Wei H., Xiang L., Wayne A. S., Chertov O., FitzGerald D. J., Bera T. K., Pastan I.. ( 2012;). Immunotoxin resistance via reversible methylation of the DPH4 promoter is a unique survival strategy. . Proc Natl Acad Sci U S A 109:, 6898–6903. [CrossRef][PubMed]
    [Google Scholar]
  72. Wellbrock C., Rana S., Paterson H., Pickersgill H., Brummelkamp T., Marais R.. ( 2008;). Oncogenic BRAF regulates melanoma proliferation through the lineage specific factor MITF. . PLoS One 3:, e2734. [CrossRef][PubMed]
    [Google Scholar]
  73. Wood S., Pithadia R., Rehman T., Zhang L., Plichta J., Radek K. A., Forsyth C., Keshavarzian A., Shafikhani S. H.. ( 2013;). Chronic alcohol exposure renders epithelial cells vulnerable to bacterial infection. . PLoS One 8:, e54646. [CrossRef][PubMed]
    [Google Scholar]
  74. Zhu S., Korzh V., Gong Z., Low B. C.. ( 2008;). RhoA prevents apoptosis during zebrafish embryogenesis through activation of Mek/Erk pathway. . Oncogene 27:, 1580–1589. [CrossRef][PubMed]
    [Google Scholar]
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