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Use of Tumor-Targeting Trans-Splicing Ribozyme for Cancer Treatment

  • Seong-Wook Lee
  • Jin-Sook Jeong
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1103)

Abstract

One of the major concerns with regard to successful cancer gene therapy is to enhance both efficacy and safety. Gene targeting may represent an attractive tool to combat cancer cells without damage to normal cells. Here, we introduce a tumor-targeting approach with the Tetrahymena group I intron-based trans-splicing ribozyme, which cleaves target RNA and trans-ligate an exon tagged at the end of the ribozyme onto the downstream U nucleotide of the cleaved target RNA. We develop a specific trans-splicing ribozyme that can target and reprogram human cytoskeleton-associate protein 2 (hCKAP2)-encoding RNA to trigger therapeutic transgene herpes simplex virus thymidine kinase (HSVtk) selectively in cancer cells that express the RNA. Adenoviral vectors encoding the hCKAP2-specific trans-splicing ribozyme are constructed for in vivo delivery into either subcutaneous tumor xenograft or orthotopically multifocal hepatocarcinoma. We present analyses of the efficacy of the recombinant adenoviral vectors in terms of cancer retardation, target RNA and cell specificity, and in vivo toxicity.

Key words

Ribozyme Trans-splicing Group I Intron RNA replacement Adenovirus Cancer gene therapy 

Notes

Acknowledgement

This work was supported by grants from National Research Foundation of Korea funded by the Ministry of Science, ICT and Future Planning (2011-0002169, 2012R1A1A2A10039116, 2012M3A9B6055200) and a grant from National R&D Program for Cancer Control, Korean Ministry of Health & Welfare (0720520).

References

  1. 1.
    McCormick F (2001) Cancer gene therapy: fringe or cutting edge? Nat Rev Cancer 1:130–141PubMedCrossRefGoogle Scholar
  2. 2.
    Wu L, Johnson M, Sato M (2003) Transcriptionally targeted gene therapy to detect and treat cancer. Trends Mol Med 9:421–429PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Tarner IH, Muller-Ladner U, Fathman CG (2004) Targeted gene therapy: frontiers in the development of ‘smart drugs’. Trends Biotechnol 22:304–310PubMedCrossRefGoogle Scholar
  4. 4.
    Been MD, Cech TR (1986) One binding site determines sequence specificity of Tetrahymena pre-rRNA self-splicing, trans-splicing, and RNA enzyme activity. Cell 47:207–216Google Scholar
  5. 5.
    Sullenger BA, Cech TR (1994) Ribozyme-mediated repair of defective mRNA by targeted, trans-splicing. Nature 371:619–622Google Scholar
  6. 6.
    Jones JT, Lee SW, Sullenger BA (1996) Tagging ribozyme reaction sites to follow trans-splicing in mammalian cells. Nat Med 2:643–648Google Scholar
  7. 7.
    Lan N, Howrey RP, Lee SW, Smith CA, Sullenger BA (1998) Ribozyme-mediated repair of sickle beta-globin mRNAs in erythrocyte precursors. Science 280:1593–1596PubMedCrossRefGoogle Scholar
  8. 8.
    Phylactou LA, Darrah C, Wood MJ (1998) Ribozyme-mediated trans-splicing of a trinucleotide repeat. Nat Genet 18:378–381Google Scholar
  9. 9.
    Lan N, Rooney BL, Lee SW, Howrey RP, Smith CA, Sullenger BA (2000) Enhancing RNA repair efficiency by combining trans-splicing ribozymes that recognize different accessible sites on a target RNA. Mol Ther 2:245–255Google Scholar
  10. 10.
    Watanabe T, Sullenger BA (2000) Induction of wild-type p53 activity in human cancer cells by ribozymes that repair mutant p53 transcripts. Proc Natl Acad Sci U S A 97:8490–8494Google Scholar
  11. 11.
    Rogers CS, Vanoye CG, Sullenger BA, George AL Jr (2002) Functional repair of a mutant chloride channel using a trans-splicing ribozyme. J Clin Invest 110:1783–1789Google Scholar
  12. 12.
    Ryu KJ, Kim JH, Lee SW (2003) Ribozyme-mediated selective induction of new gene activity in hepatitis C virus internal ribosome entry site-expressing cells by targeted trans-splicing. Mol Ther 7:386–395Google Scholar
  13. 13.
    Shin KS, Sullenger BA, Lee SW (2004) Ribozyme-mediated induction of apoptosis in human cancer cells by targeted repair of mutant p53 RNA. Mol Ther 10:365–372PubMedCrossRefGoogle Scholar
  14. 14.
    Kastanos E, Hjiantoniou E, Phylactou LA (2004) Restoration of protein synthesis in pancreatic cancer cells by trans-splicing ribozymes. Biochem Biophys Res Commun 322:930–934Google Scholar
  15. 15.
    Kwon BS, Jung HS, Song MS, Cho KS, Kim SC, Kimm K, Jeong JS, Kim IH, Lee SW (2005) Specific regression of human cancer cells by ribozyme-mediated targeted replacement of tumor-specific transcript. Mol Ther 12:824–834Google Scholar
  16. 16.
    Hong SH, Jeong JS, Lee YJ, Jung HI, Cho KS, Kim CM, Kwon BS, Sullenger BA, Lee SW, Kim IH (2008) In vivo reprogramming of hTERT by trans-splicing ribozyme to target tumor cells. Mol Ther 16:74–80Google Scholar
  17. 17.
    Jeong JS, Lee SW, Hong SH, Lee YJ, Jung HI, Cho KS, Seo HH, Lee SJ, Park S, Song MS, Kim CM, Kim IH (2008) Antitumor effects of systemically delivered adenovirus harboring trans-splicing ribozyme in intrahepatic colon cancer mouse model. Clin Cancer Res 14:281–290Google Scholar
  18. 18.
    Song MS, Jeong JS, Ban G, Lee JH, Won YS, Cho KS, Kim IH, Lee SW (2009) Validation of tissue-specific promoter-driven tumor-targeting trans-splicing ribozyme system as a multifunctional cancer gene therapy device in vivo. Cancer Gene Ther 16:113–125Google Scholar
  19. 19.
    Jung HS, Lee SW (2006) Ribozyme-mediated selective killing of cancer cells expressing carcinoembryonic antigen RNA by targeted trans-splicing. Biochem Biophys Res Commun 349:556–563PubMedCrossRefGoogle Scholar
  20. 20.
    Won YS, Lee SW (2007) Targeted retardation of hepatocarcinoma cells by specific replacement of alpha-fetoprotein RNA. J Biotechnol 129:614–619PubMedCrossRefGoogle Scholar
  21. 21.
    Won YS, Lee SW (2012) Selective regression of cancer cells expressing a splicing variant of AIMP2 through targeted RNA replacement by trans-splicing ribozyme. J Biotechnol 158:44–49PubMedCrossRefGoogle Scholar
  22. 22.
    Ban G, Jeong JS, Kim A, Kim SJ, Han SY, Kim IH, Lee SW (2011) Selective and efficient retardation of cancers expressing cytoskeleton-associated protein 2 by targeted RNA replacement. Int J Cancer 129:1018–1029PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Seong-Wook Lee
    • 1
  • Jin-Sook Jeong
    • 2
  1. 1.Department of Molecular Biology, Institute of Nanosensor and BiotechnologyDankook UniversityYonginSouth Korea
  2. 2.Department of Pathology and Medical Research Center for Cancer Molecular TherapyDong-A University College of MedicineBusanSouth Korea

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