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Therapeutic Ribozymes

Principles and Applications

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Summary

Ribozymes have progressed from an intriguing subject of scientific study to therapeutic agents for the potential treatment of both acquired and inherited diseases. Clinical trials using ribozymes targeted against HIV, the aetiological agent of AIDS, have recently been initiated. Despite this rapid progression to clinical application, there are many unexplored avenues which still must be examined to improve the intracellular effectiveness of ribozymes. Since ribozymes are RNA molecules which selectively cleave RNA targets via base-pairing interactions, the rules governing nucleic acid hybridisation affect ribozyme function. In addition, knowledge of the cellular mechanisms governing RNA partitioning and stability apply equally to ribozymes as they do to the target RNAs. The successful therapeutic application of ribozymes depends upon increasing our knowledge of RNA metabolism and movement, and applying this knowledge in the design of ribozymes. This review summarises some of the progress and experimental approaches towards achieving these goals as well as surveying experimental testing of potential therapeutic applications. It is becoming increasingly evident that ribozymes can serve the dual function of a tool to elucidate the functional roles of many gene products as well as a therapeutic agent designed to functionally destroy deleterious RNAs.

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References

  1. Krager K, Grabowski PJ, Zaug AJ, et al. Self-splicing RNA: autoexcision and autocyclization of the ribosomal RNA intervening sequence of Tetrahymena. Cell 1982; 31: 147–57

    Article  Google Scholar 

  2. Guerrier-Takada C, Gardiner K, Marsh T, et al. The RNA moiety of ribonuclease P is the catalytic subunit of the enzyme. Cell 1983; 35: 849–57

    Article  PubMed  CAS  Google Scholar 

  3. Cech TR. Ribozyme engineering. Curr Opin Struct Biol 1992; 2: 605–9

    Article  CAS  Google Scholar 

  4. Rossi JJ. Making ribozymes work in cells. Curr Biol 1994; 4: 469–71

    Article  PubMed  CAS  Google Scholar 

  5. Sullenger BA, Cech TR. Ribozyme-mediated repair of defective mRNA by targeted, trans-splicing. Nature 1992; 371: 619–22

    Article  Google Scholar 

  6. Young B, Herschlag D, Cech TR. Mutations in a nonconserved sequence of the Tetrahymena ribozyme increase activity and specificity. Cell 1991; 29: 10159–71

    Google Scholar 

  7. Dahm SC, Derrick WB, Uhlenbeck OC. Evidence for the role of solvated metal hydroxide in the hammerhead cleavage mechanism. Biochemistry 1993; 32: 13040–5

    Article  PubMed  CAS  Google Scholar 

  8. Ruffner DE, Stormo GD, Uhlenbeck OC. Sequence requirements of the hammerhead RNA self-cleavage reaction. Biochemistry 1990; 29: 10695–702

    Article  PubMed  CAS  Google Scholar 

  9. Goodchild J, Kohli V. Ribozymes that cleave an RNA sequence from human immunodeficiency virus: the effect of flanking sequence on rate. Arch Biochem Biophys 1991; 284: 386–91

    Article  PubMed  CAS  Google Scholar 

  10. Hendry P, McCall MJ, Santiago FS, et al. A ribozyme with DNA in the hybridising arms displays enhanced cleavage ability. Nucleic Acids Res 1992; 20: 5737–41

    Article  PubMed  CAS  Google Scholar 

  11. Paolella G, Sproat BS, Lamond AI. Nuclease resistant ribozymes with high catalytic activity. EMBO J 1992; 11: 1913–9

    PubMed  CAS  Google Scholar 

  12. Taylor NR, Kaplan BE, Swiderski P, et al. Chimeric DNA-RNA hammerhead ribozymes have enhanced in vitro catalytic efficiency and increased stability in vivo. Nucleic Acids Res 1992; 20: 4559–65

    Article  PubMed  CAS  Google Scholar 

  13. Bratty J, Chartrand P, Ferbeyre G, et al. The hammerhead RNA domain, a model ribozyme. Biochim Biophys Acta 1993; 1216: 345–9

    Article  PubMed  CAS  Google Scholar 

  14. Heidenreich O, Benseier P, Fahrenholz A, et al. High activity and stability of hammerhead ribozymes containing 2′-modified pyrimidine nucleosides and phosphorothioates. J Biol Chem 1994; 269: 2131–8

    PubMed  CAS  Google Scholar 

  15. Jarvis TC, Wincott FE, Laverna JA, et al. Optimizing the cell efficacy of synthetic ribozymes: site selection and chemical modifications of ribozymes targeting the proto-oncogene c-myb. J Biol Chem 1996; 271: 29107–12

    Article  PubMed  CAS  Google Scholar 

  16. Hertel KJ, Herschlag D, Uhlenbeck OC. A kinetic and thermodynamic framework for the hammerhead ribozyme reaction. Biochemistry 1994; 33: 3374–85

    Article  PubMed  CAS  Google Scholar 

  17. Hertel KJ, Uhlenbeck OC. The internal equilibrium of the hammerhead ribozyme reaction. Biochemistry 1995; 34: 1744–9

    Article  PubMed  CAS  Google Scholar 

  18. Dropulic B, Lin NH, Martin MA, et al. Functional characterization of a U5 ribozyme: intracellular suppression of human immunodeficiency virus type 1 expression. J Virol 1992; 66: 1432–41

    PubMed  CAS  Google Scholar 

  19. Dropulic B, Jeang KT. A virus model for examining the intracellular efficiency of a ribozyme targeted to HIV-1. Methods: a Companion to Methods in Enzymology 1993; 5: 43–9

    Article  CAS  Google Scholar 

  20. Xing Z, Whitton JL. An anti-lymphocytic choriomeningitis virus ribozyme expressed in tissue culture cells diminishes viral RNA levels and leads to a reduction in infectious virus yield. J Virol 1993; 67: 1840–7

    PubMed  CAS  Google Scholar 

  21. Bertrand E, Pictet R, Frange T. Can hammerhead ribozymes be efficient tools to inactivate gene function? [published erratum appears in Nucleic Acids Res 1994 Apr 11; 22 (7): 1326]. Nucleic Acids Res 1994; 22: 293–300

    Article  PubMed  CAS  Google Scholar 

  22. Sullenger B, Cech TR. Tethering ribozymes to a retroviral packaging signal for destruction of viral RNA. Science 1993; 262: 1566–9

    Article  PubMed  CAS  Google Scholar 

  23. Cameron FH, Jennings PA. Specific gene suppression by engineered ribozymes in monkey cells. Proc Natl Acad Sci USA 1989; 86: 9139–43

    Article  PubMed  CAS  Google Scholar 

  24. Steinecke P, Herget T, Schreier PH. Expression of a chimeric ribozyme gene results in endonucleolytic cleavage of target mRNA and a concomitant reduction of gene expression in vivo. EMBO J 1992; 11: 1525–30

    PubMed  CAS  Google Scholar 

  25. Zhao JJ, Pick L. Generating loss-of-function phenotypes of the fushi tarazu gene with a targeted ribozyme in Drosophila. Nature 1993; 365: 448–51

    Article  PubMed  CAS  Google Scholar 

  26. Efrat S, Fusco-DeMane M, Wu Y-J, et al. Ribozyme-mediated attenuation of pancreatic beta-cell glucokinase expression in transgenic mice results in impaired glucose-induced insulin secretion. Proc Natl Acad Sci USA 1994; 91: 2051–5

    Article  PubMed  CAS  Google Scholar 

  27. Larsson S, Hotchkiss G, Andang M, et al. Reduced β2-microglobulin mRNA levels in transgenic mice expressing a designed hammerhead ribozyme. Nucleic Acids Res 1994; 22: 2242–8

    Article  PubMed  CAS  Google Scholar 

  28. Sarver N, Cantin EM, Chang PS, et al. Ribozymes as potential anti-HIV-1 therapeutic agents. Science 1990; 247: 1222–5

    Article  PubMed  CAS  Google Scholar 

  29. Zhou C, Bahner IC, Larson GP, et al. Inhibition of HIV-1 in human T-lymphocytes by retrovirally transduced anti-tat and rev hammerhead ribozymes. Gene 1994; 149: 33–9

    Article  PubMed  CAS  Google Scholar 

  30. Bauer G, Valdez P, Kearns K, et al. Inhibition of human immunodeficiency virus-1 (HIV-1) replication after transduction of granulocyte colony-stimulating factor-mobilized CD34+ cells from HIV-1-infected donors using retroviral vectors containing anti-HIV-1 genes. Blood 1997; 89(7): 2259–67

    PubMed  CAS  Google Scholar 

  31. Chen CJ, Banerjea AC, Harmison GG, et al. Multitarget-ribozyme directed to cleave at up to nine highly conserved HIV-1 env RNA regions inhibits HIV-1 replication: potential effectiveness against most presently sequenced HIV-1 isolates. Nucleic Acids Res 1992; 20: 4581–9

    Article  PubMed  CAS  Google Scholar 

  32. Crisell P, Thompson S, James W. Inhibition of HIV-1 replication by ribozymes that show poor activity in vitro. Nucleic Acids Res 1993; 21: 5251–5

    Article  PubMed  CAS  Google Scholar 

  33. Dropulic B, Hermankova M, Pitha PM. A conditionally replicating HIV-1 vector interferes with wild-type HIV-1 replication and spread. Proc Natl Acad Sci USA 1996; 93: 11103–8

    Article  PubMed  CAS  Google Scholar 

  34. Lo KM, Biasolo MA, Dehni G, et al. Inhibition of replication of HIV-1 by retroviral vectors expressing tat-antisense and anti-tat ribozyme RNA. Virology 1992; 190: 176–83

    Article  PubMed  CAS  Google Scholar 

  35. Ojwang JO, Hampel A, Looney DJ, et al. Inhibition of human immunodeficiency virus type 1 expression by a hairpin ribozyme. Proc Natl Acad Sci USA 1992; 89: 10802–6

    Article  PubMed  CAS  Google Scholar 

  36. Weerasinghe M, Liem SE, Asad S, et al. Resistance to human immunodeficiency virus type 1 (HIV-1) infection in human CD4+ lymphocyte-derived cell lines conferred by using retroviral vectors expressing an HIV-1 RNA-specific ribozyme. J Virol 1991; 65: 5531–4

    PubMed  CAS  Google Scholar 

  37. Yamada O, Yu M, Yee JK, et al. Intracellular immunization of human T cells with a hairpin ribozyme against human immunodeficiency virus type 1. Gene Ther 1994; 1: 38–45

    PubMed  CAS  Google Scholar 

  38. Yu M, Ojwang J, Yamada O, et al. A hairpin ribozyme inhibits expression of diverse strains of human immunodeficiency virus type 1 [published erratum appears in Proc Natl Acad Sci USA 1993 Sep 1; 90 (17): 8303]. Proc Natl Acad Sci USA 1993; 90: 6340–4

    Article  PubMed  CAS  Google Scholar 

  39. Zack JA, Arrigo SJ, Weitsman SR, et al. HIV-1 entry into quiescent primary lymphocytes: molecular analysis reveals a labile, latent viral structure. Cell 1990; 61: 213–22

    Article  PubMed  CAS  Google Scholar 

  40. Lieber A, He CY, Polyak SJ, et al. Elimination of hepatitis C virus RNA in infected human hepatocytes by adenovirus-mediated expression of ribozymes. J Virol 1996; 70: 8782–91

    PubMed  CAS  Google Scholar 

  41. Koizumi M, Kamiya H, Ohtsuka E. Ribozymes designed to inhibit transformation of NIH3T3 cells by the activated c-Ha-ras gene. Gene 1992; 117: 179–84

    Article  PubMed  CAS  Google Scholar 

  42. Li M, Lonial H, Citarella R, et al. Tumor inhibitory activity of anti-ras ribozymes delivered by retroviral gene transfer. Cancer Gene Ther 1996; 3: 221–9

    PubMed  CAS  Google Scholar 

  43. Snyder DS, Wu Y, Wang JL, et al. Ribozyme-mediated inhibition of bcr-abl gene expression in a Philadelphia chromosome-positive cell line. Blood 1993; 82: 600–5

    PubMed  CAS  Google Scholar 

  44. Shore SK, Nabissa PM, Reddy ER Ribozyme-mediated cleavage of the BCR-ABL oncogene transcript: in vitro cleavage of RNA and in vivo loss of P210 protein-kinase activity. Oncogene 1993; 8: 3183–8

    PubMed  CAS  Google Scholar 

  45. Daly C, Coyle S, McBride S, et al. mdrl ribozyme mediated reversal of the multi-drug resistant phenotype in human lung cell lines. Cytotechnology 1996; 19: 199–205

    Article  PubMed  CAS  Google Scholar 

  46. Czubayko F, Schulte AM, Berchem GJ, et al. Melanoma angiogenesis and metastasis modulated by ribozyme targeting of the secreted growth factor pleiotrophin. Proc Natl Acad Sci USA 1996: 93: 14753–8

    Article  PubMed  CAS  Google Scholar 

  47. Kanazawa Y, Ohkawa K, Ueda K, et al. Hammerhead ribozyme-mediated inhibition of telomerase activity in extracts of human hepatocellular carcinoma cells. Biochem Biophys Res Commun 1996; 225: 570–6

    Article  PubMed  CAS  Google Scholar 

  48. Kilpatrick MW, Phylactou LA, Godfrey M, et al. Delivery of a hammerhead ribozyme specifically down-regulates the production of fibrillin-1 by cultured dermal fibroblasts. Hum Mol Genet 1966; 5: 1939–44

    Article  Google Scholar 

  49. Sioud M. Ribozyme modulation of lipopolysaccharide-induced tumor necrosis factor-alpha production by peritoneal cells in vitro and in vivo. Eur J Immunol 1996; 26: 1026–31

    Article  PubMed  CAS  Google Scholar 

  50. Turck J, Pollock AS, Lee LK, et al. Matrix metalloproteinase 2 (gelatinase A) regulates glomerular mesangial cell proliferation and differentiation. J Biol Chem 1996; 271: 15074–83

    Article  PubMed  CAS  Google Scholar 

  51. Jarvis TC, Alby LJ, Beaudry AA, et al. Inhibition of vascular smooth muscle cell proliferation by ribozymes that cleave c-myb mRNA. RNA 1996; 2: 419–28

    PubMed  CAS  Google Scholar 

  52. Morgan RA, Anderson WF. Human gene therapy. Annu Rev Biochem 1993; 62: 191–217

    Article  PubMed  CAS  Google Scholar 

  53. Muzcyzka N. Curr Top Microbiol Immunol 1992; 158: 7–123

    Google Scholar 

  54. Bertrand EL, Rossi JJ. Facilitation of hammerhead ribozyme catalysis by the nucleocapsid protein of HIV-1 and the heterogeneous nuclear ribonucleoprotein A1. EMBO J 1994; 13: 2904–12

    PubMed  CAS  Google Scholar 

  55. Herschlag D, Khosla M, Tsuchihashi Z, et al. An RNA chaper-one activity of non-specific RNA binding proteins in hammerhead ribozyme catalysis [published erratum appears in EMBO J 1994 Aug 15; 13(16): 3926]. EMBO J 1994; 13: 2913–24

    PubMed  CAS  Google Scholar 

  56. Tsuchihashi Z, Khoslo M, Herschlag D. Protein enhancement of hammerhead ribozyme catalysis. Science 1993; 262: 99–102

    Article  PubMed  CAS  Google Scholar 

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  1. Department of Molecular Biology, Beckman Research Institute of the City of Hope, 1450 E. Duarte Road, Duarte, California, CA 91010, USA

    John J. Rossi

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Correspondence to John J. Rossi.

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Rossi, J.J. Therapeutic Ribozymes. BioDrugs 9, 1–10 (1998). https://doi.org/10.2165/00063030-199809010-00001

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