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Adenovirus-mediated FIR demonstrated TP53-independent cell-killing effect and enhanced antitumor activity of carbon-ion beams

Gene Therapy volume 23, pages 50–56 (2016)Cite this article

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Abstract

Combination therapy of carbon-ion beam with the far upstream element-binding protein (FBP)-interacting repressor, FIR, which interferes with DNA damage repair proteins, was proposed as an approach for esophageal cancer treatment with low side effects regardless of TP53 status. In vivo therapeutic antitumor efficacy of replication-defective adenovirus (E1 and E3 deleted adenovirus serotype 5) encoding human FIR cDNA (Ad-FIR) was demonstrated in the tumor xenograft model of human esophageal squamous cancer cells, TE-2. Bleomycin (BLM) is an anticancer agent that introduces DNA breaks. The authors reported that Ad-FIR involved in the BLM-induced DNA damage repair response and thus applicable for other DNA damaging agents. To examine the effect of Ad-FIR on DNA damage repair, BLM, X-ray and carbon-ion irradiation were used as DNA damaging agents. The biological effects of high linear energy transfer (LET) radiotherapy used with carbon-ion irradiation are more expansive than low-LET conventional radiotherapy, such as X-rays or γ rays. High LET radiotherapy is suitable for the local control of tumors because of its high relative biological effectiveness. Ad-FIR enhanced BLM-induced DNA damage indicated by γH2AX in vitro. BLM treatment increased endogenous nuclear FIR expression in TE-2 cells, and P27Kip1 expression was suppressed by TP53 siRNA and BLM treatment. Further, Ad-FIRΔexon2, a dominant-negative form of FIR that lacks exon2 transcriptional repression domain, decreased Ku86 expression. The combination of Ad-FIR and BLM in TP53 siRNA increased DNA damage. Additionally, Ad-FIR showed synergistic cell toxicity with X-ray in vitro and significantly increased the antitumor efficacy of carbon-ion irradiation in the xenograft mouse model of TE-2 cells (P=0.03, Mann–Whitney’s U-test) and was synergistic with the sensitization enhancement ratio (SER) value of 1.15. Therefore, Ad-FIR increased the cell-killing activity of the carbon-ion beam that avoids late-phase severe adverse effects independently of the TP53 status in vitro. Our findings indicated the feasibility of the combination of Ad-FIR with DNA damaging agents for future esophageal cancer treatment.

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References

  1. Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM . Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer 2010; 127: 2893–2917.

    Article  CAS  Google Scholar 

  2. Mirzayans R, Andrais B, Scott A, Murray D . New insights into p53 signaling and cancer cell response to DNA damage: implications for cancer therapy. J Biomed Biotechnol 2012; 2012: 170325.

    Article  Google Scholar 

  3. Sano D, Xie TX, Ow TJ, Zhao M, Pickering CR, Zhou G et al. Disruptive TP53 mutation is associated with aggressive disease characteristics in an orthotopic murine model of oral tongue cancer. Clin Cancer Res 2011; 17: 6658–6670.

    Article  CAS  Google Scholar 

  4. Shimada H, Liu TL, Ochiai T, Shimizu T, Haupt Y, Hamada H et al. Facilitation of adenoviral wild-type p53-induced apoptotic cell death by overexpression of p33ING1 in T.Tn human esophageal carcinoma cells. Oncogene 2002; 21: 1208–1216.

    Article  CAS  Google Scholar 

  5. Shimada H, Matsubara H, Shiratori T, Shimizu T, Miyazaki S, Okazumi S et al. Phase I/II adenoviral p53 gene therapy for chemoradiation resistant advanced esophageal squamous cell carcinoma. Cancer Sci 2006; 97: 554–561.

    Article  CAS  Google Scholar 

  6. Kawahira H, Matsushita K, Shimada H, Shimizu T, Nabeya Y, Hayashi H et al. Viral shedding after p53 adenoviral gene therapy in 10 cases of esophageal cancer. Cancer Sci 2010; 101: 289–291.

    Article  CAS  Google Scholar 

  7. Liu J, Akoulitchev S, Weber A, Ge H, Chuikov S, Libutti D et al. Defective interplay of activators and repressors with TFIH in xeroderma pigmentosum. Cell 2001; 104: 353–363.

    Article  CAS  Google Scholar 

  8. Matsushita K, Tomonaga T, Shimada H, Shioya A, Higashi M, Matsubara H et al. An essential role of alternative splicing of c-myc suppressor FIR in carcinogenesis. Cancer Res 2006; 66: 1409–1417.

    Article  CAS  Google Scholar 

  9. Kajiwara T, Matsushita K, Itoga S, Tamura M, Tanaka N, Tomonaga T et al. SAP155-mediated c-myc suppressor far-upstream element-binding protein-interacting repressor splicing variants are activated in colon cancer tissues. Cancer Sci 2013; 104: 149–156.

    Article  CAS  Google Scholar 

  10. Matsushita K, Kitamura K, Rahmutulla B, Tanaka N, Ishige T, Satoh M et al. Haploinsufficiency of the c-myc transcriptional repressor FIR, as a dominant negative-alternative splicing model, promoted p53-dependent T-cell acute lymphoblastic leukemia progression by activating Notch1. Oncotarget 2014; 6: 5102–5117.

    PubMed Central  Google Scholar 

  11. Rahmutulla B, Matsushita K, Satoh M, Seimiya M, Tsuchida S, Kubo S et al. Alternative splicing of FBP-interacting repressor coordinates c-Myc, P27Kip1/cyclinE and Ku86/XRCC5 expression as a molecular sensor for bleomycin-induced DNA damage pathway. Oncotarget 2014; 5: 2404–2417.

    Article  Google Scholar 

  12. Matsushita K, Tamura M, Tanaka N, Tomonaga T, Matsubara H, Shimada H et al. Interactions between SAP155 and FUSE-binding protein-interacting repressor bridges c-myc and P27Kip1 expression. Mol Can Res 2013; 11: 689–698.

    Article  CAS  Google Scholar 

  13. Matsushita K, Kajiwara T, Tamura M, Satoh M, Tanaka N, Tomonaga T et al. SAP155-mediated splicing of FUSE-binding protein-interacting repressor (FIR) serves as a molecular switch for c-myc gene expression. Mol Cancer Res 2012; 10: 787–799.

    Article  CAS  Google Scholar 

  14. Rahmutulla B, Matsushita K, Nomura F . Alternative splicing of DNA damage response genes and gastrointestinal cancers. World J Gastroenterol 2014; 20: 17305–17313.

    Article  Google Scholar 

  15. Tanaka N, Araki K, Mizokami D, Miyagawa Y, Yamashita T, Tomifuji M et al. Sendai virus-mediated c-myc suppressor far-upstream element binding protein interacting repressor gene transfer suppresses head and neck squamous cell carcinoma. Gene Therapy 2015.

  16. Kitamura A, Matsushita K, Takiguchi Y, Shimada H, Tada Y, Yamanaka M et al. Synergistic effect of non-transmissible Sendai virus vector encoding the c-myc suppressor FUSE-binding protein-interacting repressor plus cisplatin in treatment of malignant pleural mesothelioma. Cancer Sci 2011; 7: 1366–1373.

    Article  Google Scholar 

  17. Mizokami D, Araki K, Tanaka N, Suzuki H, Tomifuji M, Yamashita T et al. Gene therapy of c-myc suppressor FUSE-binding protein-interacting repressor by Sendai virus delivery prevents tracheal stenosis. Plos One 2015; 10: e0116279.

    Article  Google Scholar 

  18. Matsushita K, Tomonaga T, Kajiwara T, Shimada H, Itoga S, Hiwasa T, Kubo S, Ochiai T, Matsubara H, Nomura F . c-myc suppressor FBP-interacting repressor for cancer diagnosis and therapy. Front Biosci 2009; 1: 3401–3408.

    Article  Google Scholar 

  19. Akutsu Y, Yasuda S, Nagata M, Izumi Y, Okazumi S, Shimada H et al. A phase I/II clinical trial of preoperative short-course carbon-ion radiotherapy for patients with squamous cell carcinoma of the esophagus. J Surg Oncol 2012; 105: 750–755.

    Article  Google Scholar 

  20. Kano M, Yamada S, Hoshino I, Murakami K, Akutsu Y, Sakata H et al. Effects of carbon-ion radiotherapy combined with a novel histone deacetylase inhibitor, cyclic hydroxamic-acid-containing peptide 31 in human esophageal squamous cell carcinoma. Anticancer Res 2009; 29: 4433–4438.

    CAS  PubMed  Google Scholar 

  21. Tsujii H, Mizoe JE, Kamada T, Baba M, Kato S, Kato H et al. Overview of clinical experiences on carbon ion radiotherapy at NIRS. Radiother Oncol 2004; 73 (suppl 2): S41–S49.

    Article  Google Scholar 

  22. Amornwichet N, Oike T, Shibata A, Ogiwara H, Tsuchiya N, Yamauchi M et al. Carbon-ion beam irradiation kills X-ray-resistant TP53-null cancer cells by inducing mitotic catastrophe. PLoS One 2014; 9: e115121.

    Article  Google Scholar 

  23. Loeffler JS, Durante M . Charged particle therapy—optimization, challenges and future directions. Nat Rev Clin Oncol 2013; 10: 411–424.

    Article  Google Scholar 

  24. Yamada S . Postoperative Recurrence of Rectal Cancer. Carbon-Ion Radiotherapy: Principles, Practices, and Treatment Planning. Springer: Tokyo, 2014: 203–209.

    Book  Google Scholar 

  25. Matsushita K, Ochiai T, Shimada H, Kato S, Ohno T, Nikaido T et al. The effects of carbon ion irradiation revealed by excised perforated intestines as a late morbidity for uterine cancer treatment. Surg Today 2006; 36: 692–700.

    Article  Google Scholar 

  26. Matsushita K, Shimada H, Ueda Y, Inoue M, Hasegawa M, Tomonaga T et al. Non-transmissible Sendai virus vector encoding c-myc suppressor FBP-interacting repressor for cancer therapy. World J Gastroenterol 2014; 20: 4316–4328.

    Article  CAS  Google Scholar 

  27. Dong F, Soubeyrand S, Haché RJ . Activation of PARP-1 in response to bleomycin depends on the Ku antigen and protein phosphatase 5. Oncogene 2010; 29: 2093–2103.

    Article  CAS  Google Scholar 

  28. Povirk LF . DNA damage and mutagenesis by radiomimetic DNA-cleaving agents: bleomycin, neocarzinostatin and other enediynes. Mutat Res 1996; 355: 71–89.

    Article  Google Scholar 

  29. Benítez-Bribiesca L, Sánchez-Suárez P . Oxidative damage, bleomycin, and gamma radiation induce different types of DNA strand breaks in normal lymphocytes and thymocytes. A comet assay study. Ann NY Acad Sci 1999; 887: 133–149.

    Article  Google Scholar 

  30. Hoeijmakers JH . Genome maintenance mechanisms for preventing cancer. Nature 2001; 411: 366–374.

    Article  CAS  Google Scholar 

  31. Oohira G, Yamada S, Ochiai T, Matsubara H, Okazumi S, Ando K et al. Growth suppression of esophageal squamous cell carcinoma induced by heavy carbon-ion beams combined with p53 gene transfer. Int J Oncol 2004; 25: 563–569.

    CAS  PubMed  Google Scholar 

  32. Imamura M, Sawada S, Kasahara-Imamura M, Harima K, Harada K . Synergistic cell-killing effect of a combination of hyperthermia and heavy ion beam irradiation: in expectation of a breakthrough in the treatment of refractory cancers (review). Int J Mol Med 2002; 9: 11–18.

    CAS  PubMed  Google Scholar 

  33. Asakawa I, Yoshimura H, Takahashi A, Ohnishi K, Nakagawa H, Ota I et al. Radiation-induced growth inhibition in transplanted human tongue carcinomas with different P53 gene status. Anticancer Res 2002; 22: 2037–2043.

    CAS  PubMed  Google Scholar 

  34. Iwadate Y, Mizoe J, Osaka Y, Ohnishi K, Nakagawa H, Ota I et al. High linear energy transfer carbon radiation effectively kills cultured glioma cells with either mutant or wild-type p53. Int J Radiat Oncol Biol Phys 2001; 50: 803–808.

    Article  CAS  Google Scholar 

  35. Kamada T, Tsujii H, Tsuji H, Yanagi T, Mizoe JE, Miyamoto T et al. Efficacy and safety of carbon ion radiotherapy in bone and soft tissue sarcomas. Working Group for the Bone and Soft Tissue Sarcomas. J Clin Oncol 2002; 20: 4466–4471.

    Article  CAS  Google Scholar 

  36. Kadono K, Homma T, Kamahara K, Nakayama M, Satoh H, Sekizawa K et al. Effect of heavy-ion radiotherapy on pulmonary function in stage I non-small cell lung cancer patients. Chest 2002; 122: 1925–1932.

    Article  CAS  Google Scholar 

  37. Nakano T, Suzuki M, Abe A, Suzuki Y, Morita S, Mizoe J et al. The phase I/II clinical study of carbon ion therapy for cancer of the uterine cervix. Cancer J Sci Am 1999; 5: 362–369.

    CAS  PubMed  Google Scholar 

  38. Takahashi A, Mori E, Ohnishi T . The foci of DNA double strand break-recognition proteins localize with γH2AX after heat treatment. J Radiat Res (Tokyo) 2010; 51: 91–95.

    Article  CAS  Google Scholar 

  39. Takahashi K, Kanazawa H, Chan H . A case of esophageal carcinoma metastatic to the mandible and characterization of two cell lines (T.T. and T.Tn). Jpn. J Oral Maxillofac Surg 1990; 36: 307–316.

    Article  Google Scholar 

  40. Nishihira T, Kasai M, Mori S et al. Characteristics of two cell lines (TE-1 and TE-2) derived from human squamous cell carcinoma of the esophagus. Gann 1979; 70: 575–584.

    CAS  PubMed  Google Scholar 

  41. Saeki H, Kitao H, Yoshinaga K, Nakanoko T, Kubo N, Kakeji Y et al. Copy-neutral loss of heterozygosity at the p53 locus in carcinogenesis of esophageal squamous cell carcinomas associated with p53 mutations. Clin Cancer Res 2011; 17: 1731–1740.

    Article  CAS  Google Scholar 

  42. Hsu IC, Tokiwa T, Bennett W, Metcalf RA, Welsh JA, Sun T et al. p53 gene mutation and integrated hepatitis B viral DNA sequences in human liver cancer cell lines. Carcinogenesis 1993; 14: 987–992.

    Article  CAS  Google Scholar 

  43. Kimura K, Nozaki N, Enomoto T, Tanaka M, Kikuchi A . Analysis of M phase-specific phosphorylation of DNA topoisomerase II. J Biol Chem 1996; 271: 21439–21445.

    Article  CAS  Google Scholar 

  44. Hiwasa T, Shimada H, Sakaida T, Kitagawa M, Kuroiwa N, Ochiai T et al. Drug-sensitivity pattern analysis of study of functional relationship between gene products. FEBS Lett 2003; 552: 177–183.

    Article  CAS  Google Scholar 

  45. Takahashi A, Kubo M, Ma H, Nakagawa A, Yoshida Y, Isono M et al. Non-homologous end-joining repair plays a more important role than homologous recombination repair in defining radiosensitivity after exposure to high-LET radiation. Radiat Res 2014; 182: 338–344.

    Article  Google Scholar 

  46. Kanai T, Endo M, Minohara S, Miyahara N, Koyama-ito H, Tomura H et al. Biophysical characteristics of HIMAC clinical irradiation system for heavy-ion radiation therapy. Int J Radiat Oncol Biol Phys 1999; 44: 201–210.

    Article  CAS  Google Scholar 

  47. Ando K, Koike S, Nojima K, Chen YJ, Ohira C, Ando S et al. Mouse skin reactions following fractionated irradiation with carbon ions. Int J Radiat Biol 1998; 74: 129–138.

    Article  CAS  Google Scholar 

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Acknowledgements

This work is supported in part by the 21st Century COE (Center Of Excellence) Programs to Dr Takenori Ochiai from the Ministry of Education, Science, Sports and Culture of Japan. This work was supported in part by a Grant-in-Aid 18591453 to KM from the Ministry of Education, Science, Sports and Culture of Japan.

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Authors and Affiliations

  1. Department of Frontier Surgery, Graduate School of Medicine, Chiba University, Chiba City, Japan

    M Kano & H Matsubara

  2. Division of Clinical Genetics and Proteomics, Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba City, Japan

    K Matsushita, B Rahmutulla & F Nomura

  3. Center Hospital, Research Center for Charged Particle Therapy, National Institute of Radiological Sciences (NIRS), Chiba City, Japan

    S Yamada

  4. Department of Surgery, School of Medicine, Toho University, Tokyo, Japan

    H Shimada

  5. Department of Genetics, Hyogo College of Medicine, Hyogo, Japan

    S Kubo

  6. Department of Biochemistry and Genetics, Chiba University, Graduate School of Medicine, Chuo-ku, Chiba, Japan

    T Hiwasa

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Kano, M., Matsushita, K., Rahmutulla, B. et al. Adenovirus-mediated FIR demonstrated TP53-independent cell-killing effect and enhanced antitumor activity of carbon-ion beams. Gene Ther 23, 50–56 (2016). https://doi.org/10.1038/gt.2015.84

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