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Sapag A, Irrazábal T, Lobos-González L, et al. Hairpin Ribozyme Genes Curtail Alcohol Drinking: from Rational Design to in vivo Effects in the Rat. Mol Ther Nucleic Acids. 2016;5(7):e335doi: 10.1038/mtna.2016.41.
Sapag, A., Irrazábal, T., Lobos-González, L., Muñoz-Brauning, C. R., Quintanilla, M. E., & Tampier, L. (2016). Hairpin Ribozyme Genes Curtail Alcohol Drinking: from Rational Design to in vivo Effects in the Rat. Molecular therapy. Nucleic acids, 5(7), e335. https://doi.org/10.1038/mtna.2016.41
Sapag, Amalia, et al. "Hairpin Ribozyme Genes Curtail Alcohol Drinking: from Rational Design to in vivo Effects in the Rat." Molecular therapy. Nucleic acids vol. 5,7 (2016): e335. doi: https://doi.org/10.1038/mtna.2016.41
Sapag A, Irrazábal T, Lobos-González L, Muñoz-Brauning CR, Quintanilla ME, Tampier L. Hairpin Ribozyme Genes Curtail Alcohol Drinking: from Rational Design to in vivo Effects in the Rat. Mol Ther Nucleic Acids. 2016 Jul 12;5(7):e335. doi: 10.1038/mtna.2016.41. PMID: 27404720; PMCID: PMC5330938.
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Mol Ther Nucleic Acids. 2016 Jul 12;5(7):e335. doi: 10.1038/mtna.2016.41.

Hairpin Ribozyme Genes Curtail Alcohol Drinking: from Rational Design to in vivo Effects in the Rat.

Molecular therapy. Nucleic acids

Amalia Sapag, Thergiory Irrazábal, Lorena Lobos-González, Carlos R Muñoz-Brauning, María Elena Quintanilla, Lutske Tampier

Affiliations

  1. Laboratory of Gene Pharmacotherapy, Department of Pharmacological and Toxicological Chemistry, Faculty of Chemical and Pharmaceutical Sciences, Universidad de Chile, Santiago, Chile.
  2. Molecular and Clinical Pharmacology Programme, Faculty of Medicine, Universidad de Chile, Santiago, Chile.

PMID: 27404720 PMCID: PMC5330938 DOI: 10.1038/mtna.2016.41

Abstract

Ribozyme genes were designed to reduce voluntary alcohol drinking in a rat model of alcohol dependence. Acetaldehyde generated from alcohol in the liver is metabolized by the mitochondrial aldehyde dehydrogenase (ALDH2) such that diminishing ALDH2 activity leads to the aversive effects of blood acetaldehyde upon alcohol intake. A stepwise approach was followed to design genes encoding ribozymes targeted to the rat ALDH2 mRNA. In vitro studies of accessibility to oligonucleotides identified suitable target sites in the mRNA, one of which fulfilled hammerhead and hairpin ribozyme requirements (CGGUC). Ribozyme genes delivered in plasmid constructs were tested in rat cells in culture. While the hairpin ribozyme reduced ALDH2 activity 56% by cleavage and blockade (P < 0.0001), the hammerhead ribozyme elicited minor effects by blockade. The hairpin ribozyme was tested in vivo by adenoviral gene delivery to UChB alcohol drinker rats. Ethanol intake was curtailed 47% for 34 days (P < 0.0001), while blood acetaldehyde more than doubled upon ethanol administration and ALDH2 activity dropped 25% in liver homogenates, not affecting other ALDH isoforms. Thus, hairpin ribozymes targeted to 16 nt in the ALDH2 mRNA provide durable and specific effects in vivo, representing an improvement on previous work and encouraging development of gene therapy for alcoholism.

References

  1. Lancet Oncol. 2009 Feb;10(2):173-80 - PubMed
  2. Proc Natl Acad Sci U S A. 2010 May 11;107(19):8531-6 - PubMed
  3. Am J Physiol Endocrinol Metab. 2007 Aug;293(2):E531-7 - PubMed
  4. J Clin Invest. 1989 Jan;83(1):314-6 - PubMed
  5. Gene Ther. 2000 Dec;7(23):2041-50 - PubMed
  6. Nat Protoc. 2007;2(5):1236-47 - PubMed
  7. Alcohol Res. 2015;37(1):15-28 - PubMed
  8. Carcinogenesis. 2007 Apr;28(4):865-74 - PubMed
  9. Methods Cell Biol. 1994;43 Pt A:161-89 - PubMed
  10. FASEB J. 1993 Jan;7(1):25-30 - PubMed
  11. Cell. 1987 Jul 3;50(1):9-16 - PubMed
  12. Addict Biol. 2006 Sep;11(3-4):310-23 - PubMed
  13. Annu Rev Biochem. 1992;61:641-71 - PubMed
  14. Gene Ther. 2003 Aug;10(15):1258-67 - PubMed
  15. J Exp Med. 2001 Sep 3;194(5):571-80 - PubMed
  16. Acta Psychiatr Scand Suppl. 1992;369:59-65; discussion 65-6 - PubMed
  17. Mol Ther Nucleic Acids. 2015 Sep 15;4:e252 - PubMed
  18. Nature. 2002 Jul 11;418(6894):222-8 - PubMed
  19. J Virol. 1996 Nov;70(11):7498-509 - PubMed
  20. Angew Chem Int Ed Engl. 2003 May 30;42(21):2424-7 - PubMed
  21. Carcinogenesis. 2002 Nov;23 (11):1851-9 - PubMed
  22. Biochemistry. 1996 Apr 9;35(14):4445-56 - PubMed
  23. Chem Biol. 2012 Jan 27;19(1):60-71 - PubMed
  24. FEBS J. 2005 Sep;272(17):4464-74 - PubMed
  25. Biochemistry. 2003 Mar 4;42(8):2259-65 - PubMed
  26. Antisense Nucleic Acid Drug Dev. 1999 Aug;9(4):341-9 - PubMed
  27. Methods Mol Biol. 1997;74:37-43 - PubMed
  28. Chembiochem. 2003 Oct 6;4(10):991-7 - PubMed
  29. RNA. 1997 Sep;3(9):961-73 - PubMed
  30. J Stud Alcohol. 2000 Jan;61(1):5-12 - PubMed
  31. Front Behav Neurosci. 2013 Jul 08;7:80 - PubMed
  32. J Biol Chem. 1973 Jul 10;248(13):4811-9 - PubMed
  33. J Mol Biol. 2005 Feb 4;345(5):987-1001 - PubMed
  34. Am J Hum Genet. 1999 Sep;65(3):795-807 - PubMed
  35. J Biol Chem. 2001 May 4;276(18):15378-85 - PubMed
  36. Biochem Genet. 1996 Apr;34(3-4):109-16 - PubMed
  37. RNA. 2008 Apr;14(4):605-15 - PubMed
  38. Oligonucleotides. 2004 Spring;14(1):11-21 - PubMed
  39. Proc Natl Acad Sci U S A. 1998 Mar 3;95(5):2509-14 - PubMed
  40. J Mol Biol. 1990 Oct 5;215(3):403-10 - PubMed
  41. Nucleic Acids Res. 1990 Jan 25;18(2):299-304 - PubMed
  42. J Immunol. 2007 Feb 1;178(3):1767-73 - PubMed
  43. Eur J Biochem. 1997 Apr 1;245(1):1-16 - PubMed
  44. Prog Nucleic Acid Res Mol Biol. 1998;58:1-39 - PubMed
  45. Biochemistry. 1995 Mar 21;34(11):3649-54 - PubMed
  46. Nucleic Acids Res. 2005 Feb 18;33(3):1058-68 - PubMed
  47. Biochemistry. 2003 Oct 7;42(39):11427-33 - PubMed
  48. Annu Rev Pharmacol Toxicol. 2010;50:259-93 - PubMed
  49. Pharmacogenet Genomics. 2005 Jun;15(6):427-31 - PubMed
  50. J Mol Med (Berl). 1995 Feb;73(2):65-71 - PubMed
  51. Adv Exp Med Biol. 2015;848:97-116 - PubMed
  52. Antisense Nucleic Acid Drug Dev. 2002 Aug;12(4):283-8 - PubMed
  53. Life Sci. 2005 Aug 5;77(12):1331-40 - PubMed
  54. Genes Dev. 1997 Nov 1;11(21):2755-66 - PubMed
  55. Nature. 1987 Aug 13-19;328(6131):596-600 - PubMed
  56. Proc Natl Acad Sci U S A. 2007 Aug 7;104(32):12982-7 - PubMed
  57. Alcohol Clin Exp Res. 2008 Jan;32(1):52-7 - PubMed
  58. RNA. 1997 Jan;3(1):75-88 - PubMed
  59. Mol Ther. 2003 May;7(5 Pt 1):670-80 - PubMed

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