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Construction of random tumor transcriptome expression library for creating and selecting novel tumor antigens

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Tumor Biology

Abstract

Novel tumor antigens are necessary for the development of efficient tumor vaccines for overcoming the immunotolerance and immunosuppression induced by tumors. Here, we developed a novel strategy to create tumor antigens by construction of random tumor transcriptome expression library (RTTEL). The complementary DNA (cDNA) from S180 sarcoma was used as template for arbitrarily amplifying gene fragments with random primers by PCR, then ligated to the C-terminal of HSP65 in a plasmid pET28a-HSP for constructing RTTEL in Escherichia coli. A novel antigen of A5 was selected from RTTEL with the strongest immunotherapeutic effects on S180 sarcoma. Adoptive immunotherapy with anti-A5 sera also inhibited tumor growth, further confirming the key antitumor roles of A5-specific antibodies in mice. A5 contains a sequence similar to protein-L-isoaspartate (D-aspartate) O-methyltransferase (PCMT1). The antisera of A5 were verified to cross-react with PCMT1 by Western blotting assay and vice versa. Both anti-A5 sera and anti-PCMT1 sera could induce antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity toward S180 cells by in vitro assay. Further assay with fluorescent staining showed that PCMT1 is detectable on the surface of S180 cells. Summary, the strategy to construct RTTEL is potential for creating and screening novel tumor antigens to develop efficient tumor vaccines. By RTTEL, we successfully created a protein antigen of A5 with significant immunotherapeutic effects on S180 sarcoma by induction of antibodies targeting for PCMT1.

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References

  1. Buonaguro L, Petrizzo A, Tornesello ML, Buonaguro FM. Translating tumor antigens into cancer vaccines. Clin Vaccine Immunol. 2011;18(1):23–34.

    Article  CAS  PubMed  Google Scholar 

  2. Kiyamova R, Kostianets O, Malyuchik S, Filonenko V, Usenko V, Gurtovyy V, Khozayenko Y, Antonuk S, Old L, Gout I. Identification of tumor-associated antigens from medullary breast carcinoma by a modified SEREX approach. Mol Biotechnol. 2010;46:105–12.

    Article  CAS  PubMed  Google Scholar 

  3. Kempa TJ, Hildesheimb A, Safaeianb M, Daunera JG, Pana Y, Porrasc C, Schillerd JT, Lowyd DR, Herreroc R, Pinto LA. HPV16/18 L1 VLP vaccine induces cross-neutralizing antibodies that may mediate cross-protection. Vaccine. 2011;29(11):2011–4.

  4. Kruit WH, Suciu S, Dreno B, Mortier L, Robert C, Chiarion-Sileni V, Maio M, Testori A, Dorval T, Grob J, Becker JC, Spatz A, Eggermont AM, Louahed J, Lehmann FF, Brichard VG, Keilholz U. Selection of immunostimulant AS15 for active immunization with MAGE-A3 protein: results of a randomized phase II study of the European Organisation for Research and Treatment of Cancer melanoma Group in Metastatic Melanoma. J Clin Oncol. 2013;31(19):2413–20.

    Article  CAS  PubMed  Google Scholar 

  5. Vansteenkiste J, Zielinski M, Linder A, Dahabreh J, Gonzalez EE, Malinowski W, Lopez-Brea M, Vanakesa T, Jassem J, Kalofonos H, Perdeus J, Bonnet R, Basko J, Janilionis R, Passlick B, Treasure T, Gillet M, Lehmann FF, Brichard VG. Adjuvant MAGE-A3 immunotherapy in resected non-small-cell lung cancer: phase II randomized study results. J Clin Oncol. 2013;31(19):2396–403.

    Article  CAS  PubMed  Google Scholar 

  6. Rosenberg SA. The curative potential of human cancer immunotherapy. Sci Transl Med. 2012;4:1–7.

    Article  Google Scholar 

  7. Thomas AA, Fisher JL, Ernstoff MS, Fadul CE. Vaccine-based immunotherapy for glioblastoma. CNS Oncol. 2013;2(4):331–49.

    Article  CAS  PubMed  Google Scholar 

  8. Parmiani G, De Filippo A, Novellino L, Castelli C. Unique human tumor antigens: immunobiology and use in clinical trials. J Immunol. 2007;178:1975–9.

    Article  CAS  PubMed  Google Scholar 

  9. Kim KM, Song MH, Kim MJ, Daudi S, Miliotto A, Old L, Odunsi K, Lee SY. A novel cancer/testis antigen KP-OVA-52 identifid by SEREX in human ovarian cancer is regulated by DNA methylation. Int J Oncol. 2012;41:1139–47.

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Song MH, Choi KU, Shin DH, Lee CH, Lee SY. Identification of the cancer/testis antigens AKAP3 and CTp11 by SEREX in hepatocellular carcinoma. Oncol Rep. 2012;28:1792–8.

    CAS  PubMed  Google Scholar 

  11. Yang HS, Zhang DM, Deng HX, Peng F, Wei YQ. Antitumor and anti-angiogenesis immunity induced by CR-SEREX-identified Xenopus RHAMM. Cancer Sci. 2010;101:862–8.

    Article  CAS  PubMed  Google Scholar 

  12. Cooper CD, Lawrie CH, Liggins AP, Collins GP, Hatton CS, Pulford K, Banham AH. Identification and characterization of peripheral T-cell lymphoma-associated SEREX antigens. PLoS One. 2011;6(8):e23916.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Song MH, Ha JC, Lee SM, Park YM, Lee SY. Identification of BCP-20 (FBXO39) as a cancer/testis antigen from colon cancer patients by SEREX. Biochem Biophys Res Commun 2011. 2011;408:195–201.

    Article  CAS  Google Scholar 

  14. Clark LC, Combs Jr GF, Turnbull BW, Slate EH, Chalker DK, Chow J, Davis LS, Glover RA, Graham GF, Gross EG, Krongrad A, Lesher Jr JL, Park HK, Sanders Jr BB, Smith CL, Taylor JR. Effects of selenium supplementation for cancer prevention in patients with carcinoma of the skin: a randomized controlled trial. Nutritional prevention of cancer study group. JAMA. 1996;276(24):1957–63.

    Article  CAS  PubMed  Google Scholar 

  15. Nadiminty N, Gao AC. Mechanisms of selenium chemoprevention and therapy in prostate cancer. Mol Nutr Food Res. 2008;52(11):1247–60.

    Article  CAS  PubMed  Google Scholar 

  16. Chapman M, Warren EHI, Wu CJ. Applications of next-generation sequencing to blood and marrow transplantation. Biol Blood Marrow Transplant. 2012;18:S151–60.

    Article  PubMed  PubMed Central  Google Scholar 

  17. Garraway L, Lander ES. Lessons from the cancer genome. Cell. 2013;153(1):17–37.

    Article  CAS  PubMed  Google Scholar 

  18. Li L, Goedegebuure P, Mardis ER, Ellis MJ, Zhang X, Herndon JM, Fleming TP, Carreno BM, Hansen TH, Gillanders WE. Cancer genome sequencing and its pmplications for personalized cancer vaccines. Cancers. 2011;3(4):4191–211.

    Article  PubMed  PubMed Central  Google Scholar 

  19. Heubeck B, Wendler O, Bumm K, Schäfer R, Müller-Vogt U, Häusler M, Meese E, Iro H, Steinhart H. Tumor-associated antigenic pattern in squamous cell carcinomas of the head and neck-analysed by SEREX. Eur J Cancer. 2013;49:e1–7.

    Article  PubMed  Google Scholar 

  20. Rio DC, Ares Jr M, Hannon GJ, Nilsen TW. Purification of RNA using TRIzol (TRI reagent). Cold Spring Harb Protoc. 2010;6:pdb. prot5439.

    Article  Google Scholar 

  21. Mansour FH, Pestov DG. Separation of long RNA by agarose-formaldehyde gel electrophoresis. Anal Biochem. 2013;441(1):18–20.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Chandler DP, Schreckhis RW, Smith JL, Bolton Jr H. Electroelution to remove humic compounds from soil DNA and RNA extracts. J Microbiol Methods. 1997;28(1):11–9.

    Article  CAS  Google Scholar 

  23. Spandidos A, Wang X, Wang H, Seed B. PrimerBank: a resource of human and mouse PCR primer pairs for gene expression detection and quantification. Nucleic Acids Res. 2010;38(suppl 1):D792–9.

    Article  CAS  PubMed  Google Scholar 

  24. Mayr UB, Kudela P, Atrasheuskaya A, Bukin E, Ignatyev G, Lubitz W. Rectal single dose immunization of mice with Escherichia coli O157: H7 bacterial ghosts induces efficient humoral and cellular immune responses and protects against the lethal heterologous challenge. Microb Biotechnol. 2012;5(2):283–94.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Tumban E, Peabody J, Peabody DS, Chackerian B. A universal virus-like particle-based vaccine for human papillomavirus: longevity of protection and role of endogenous and exogenous adjuvants. Vaccine. 2013;31(41):4647–54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Guojun W, Wei G, Kedong O, Yi H, Yanfei X, Qingmei C, Yankai Z, Jie W, Hao F, Taiming L, Jingjing L, Rongyue C. A novel vaccine targeting gastrin-releasing peptide: efficient inhibition of breast cancer growth in vivo. Endocr Relat Cancer. 2008;15:149–59.

    Article  PubMed  Google Scholar 

  27. Jinshu X, Jingjing L, Duan P, Zheng Z, Ding M, Jie W, Rongyue C, Zhuoyi H, Roque RS. A synthetic gonadotropin-releasing hormone (GnRH) vaccine for control of fertility and hormone dependent diseases without any adjuvant. Vaccine. 2005;23:4834–43.

    Article  PubMed  Google Scholar 

  28. Sun X, Liu Y, Di D, Wu G, Guo H. Chemometric analysis of metabolism disorders in blood plasma of S180 and H22 tumor-bearing mice by high performance liquid chromatography-diode array detection. J Chemom. 2011;25(8):430–40.

    CAS  Google Scholar 

  29. Mizukami S, Kajiwara C, Ishikawa H, Katayama I, Yui K, Udono H. Both CD4 and CD8+ T cell epitopes fused to heat shock cognate protein 70 (hsc70) can function to eradicate tumors. Cancer Sci. 2008;99:1008–15.

    Article  CAS  PubMed  Google Scholar 

  30. Zhou X, Kong N, Wang J, Fan H, Zou H, Horwitz D, Brand D, Liu Z, Zheng SG. All-trans retinoic acid sustains the stability and function of natural regulatory T cells in an inflammatory milieu. J Immunol. 2010;185:2675–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Xie Y, Sun H, Li D. Platycodin D is a potent adjuvant of specific cellular and humoral immune responses against recombinant hepatitis B antigen. Vaccine. 2009;27:757–64.

    Article  CAS  PubMed  Google Scholar 

  32. Aikemu A, Umar A, Yusup A, Upur H, Berké B, Bégaud B, Bégaud B, Moore N. Immunomodulatory and antitumour effects of abnormal savda munziq on S180 tumor-bearing mice. BMC Complement Altern Med. 2012;12(1):157.

    Article  PubMed  PubMed Central  Google Scholar 

  33. Park SY, Kim YH, Kim EK, Ryu EY, Lee SJ. Heme oxygenase-1 signals are involved in preferential inhibition of pro-inflammatory cytokine release by surfactin in cells activated with Porphyromonas gingivalis lipopolysaccharide. Chem Biol Interact. 2010;188:437–45.

    Article  CAS  PubMed  Google Scholar 

  34. Nowak AK, Lake RA, Robinson BW. Combined chemoimmunotherapy of solid tumours: improving vaccines? Adv Drug Deliv Rev. 2006;58(8):975–90.

    Article  CAS  PubMed  Google Scholar 

  35. Berinstein NL. Enhancing cancer vaccines with immunomodulators. Vaccine. 2007;25s:b72–88.

    Article  Google Scholar 

  36. Gulley JL, Drake CG. Immunotherapy for prostate cancer: recent advances, lessons learned, and areas for further research. Clin Cancer Res. 2011;17(12):3884–91.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Kaufman HL, Kim-Schulze S, Manson K, DeRaffele G, Mitcham J, Seo KS, Kim DW, Marshall J. Poxvirus-based vaccine therapy for patients with advanced pancreatic cancer. J Transl Med. 2007;5:60.

    Article  PubMed  PubMed Central  Google Scholar 

  38. Sambri I, Capasso R, Pucci P, Perna AF, Ingrosso D. The microRNA 15a/16-1 cluster down-regulates protein repair isoaspartyl methyltransferase in hepatoma cells: implications for apoptosis regulation. J Biol Chem. 2011;286(51):43690–700.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Amer M, Elhefnawi M, El-Ahwany E, Awad AF, Gawad NA, Zada S, Tawab FM. Hsa-miR-195 targets PCMT1 in hepatocellular carcinoma that increases tumor life span. Tumour Biol. 2014;35(11):11301–9.

    Article  CAS  PubMed  Google Scholar 

  40. Ferris RL, Jaffee EM, Ferrone S. Tumor antigen-targeted, monoclonal antibody-based immunotherapy: clinical response, cellular immunity, and immunoescape. J Clin Oncol. 2010;28:4390–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

The authors are thankful for the project SRF150 supported by National Natural Science Foundation of China.

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Authors and Affiliations

  1. College of Life Science and Technology, Huazhong Agricultural University, No. 1 Shizishan Street, Hongshan District, Wuhan, 430070, HuBei Province, China

    Huizhun Zhao, Xiuyun Zhao & Gaofu Qi

  2. College of Life Science, Hubei University, 430062, Wuhan, China

    Peng Du

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  1. Huizhun Zhao
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Correspondence to Gaofu Qi.

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Zhao, H., Zhao, X., Du, P. et al. Construction of random tumor transcriptome expression library for creating and selecting novel tumor antigens. Tumor Biol. 37, 12877–12887 (2016). https://doi.org/10.1007/s13277-016-5201-0

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  • DOI: https://doi.org/10.1007/s13277-016-5201-0

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