Home LiteratureArticle Details
PMID: 16019264 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Intrastriatal rAAV-mediated delivery of anti-huntingtin shRNAs induces partial reversal of disease progression in R6/1 Huntington's disease transgenic mice.

Molecular therapy : the journal of the American Society of Gene Therapy ·Vol. 12 ·No. 4 ·2005-10-00 ·Pages 618-33

Rodriguez-Lebron E, Denovan-Wright EM, Nash K, Lewin AS, Mandel RJ

Abstract

Huntington's disease (HD) is a fatal neurodegenerative disorder caused by the presence of an abnormally expanded polyglutamine domain in the N-terminus of huntingtin. We developed a recombinant adeno-associated viral serotype 5 (rAAV5) gene transfer strategy to posttranscriptionally suppress the levels of striatal mutant huntingtin (mHtt) in the R6/1 HD transgenic mouse via RNA interference. Transient cotransfection of HEK293 cells with plasmids expressing a portion of human mHtt derived from R6/1 transgenic HD mice and a short-hairpin RNA directed against the 5' UTR of the mHtt mRNA (siHUNT-1) resulted in reduction in the levels of mHtt mRNA (-75%) and protein (-60%). Long-term in vivo rAAV5-mediated expression of siHUNT-1 in the striatum of R6/1 mice reduced the levels of mHtt mRNA (-78%) and protein (-28%) as determined by quantitative RT-PCR and Western blot analysis, respectively. The reduction in mHtt was concomitant with a reduction in the size and number of neuronal intranuclear inclusions and a small but significant normalization of the steady-state levels of preproenkephalin and dopamine- and cAMP-responsive phosphoprotein 32 kDa mRNA. Finally, bilateral expression of rAAV5-siHUNT-1 resulted in delayed onset of the rear paw clasping phenotype exhibited by the R6/1 mice. These results suggest that a reduction in the levels of striatal mHtt can ameliorate the HD phenotype of R6/1 mice.

MeSH Terms
Animals Dependovirus/genetics Disease Models, Animal Disease Progression Gene Expression Genetic Therapy Genetic Vectors Humans Huntingtin Protein Huntington Disease/genetics,therapy Intranuclear Inclusion Bodies Mice Mice, Transgenic/genetics Nerve Tissue Proteins/genetics,metabolism Nuclear Proteins/genetics,metabolism Phenotype Plasmids/genetics RNA Interference RNA, Messenger/metabolism RNA, Small Interfering/genetics,metabolism
Chemicals
Htt protein, mouse Huntingtin Protein Nerve Tissue Proteins Nuclear Proteins RNA, Messenger RNA, Small Interfering
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Rodriguez-Lebron Edgardo
Department of Neuroscience, University of Florida McKnight Brain Institute, Gainesville, 32610-0244, USA.
Denovan-Wright Eileen M
Nash Kevin
Lewin Alfred S
Mandel Ronald J
References (69)
69 references, click to expand
  1. Cyclic AMP-dependent activation of the proenkephalin gene requires phosphorylation of CREB at serine-133 and a Src-related kinase.
    J Neurochem. 1999 Jul;73(1):129-38 PMID: 10386963
  2. Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield.
    Gene Ther. 1999 Jun;6(6):973-85 PMID: 10455399
  3. Structure, expression and regulation of the cannabinoid receptor gene (CB1) in Huntington's disease transgenic mice.
    Eur J Biochem. 2004 Dec;271(23-24):4909-20 PMID: 15606779
  4. Mutant huntingtin affects the rate of transcription of striatum-specific isoforms of phosphodiesterase 10A.
    Eur J Neurosci. 2004 Dec;20(12):3351-63 PMID: 15610167
  5. Potent inhibition of huntingtin aggregation and cytotoxicity by a disulfide bond-free single-domain intracellular antibody.
    Proc Natl Acad Sci U S A. 2004 Dec 21;101(51):17616-21 PMID: 15598740
  6. Lentiviral-mediated silencing of SOD1 through RNA interference retards disease onset and progression in a mouse model of ALS.
    Nat Med. 2005 Apr;11(4):423-8 PMID: 15768028
  7. Silencing mutant SOD1 using RNAi protects against neurodegeneration and extends survival in an ALS model.
    Nat Med. 2005 Apr;11(4):429-33 PMID: 15768029
  8. A human single-chain Fv intrabody preferentially targets amino-terminal Huntingtin's fragments in striatal models of Huntington's disease.
    Neurobiol Dis. 2005 Jun-Jul;19(1-2):47-56 PMID: 15837560
  9. RNA interference improves motor and neuropathological abnormalities in a Huntington's disease mouse model.
    Proc Natl Acad Sci U S A. 2005 Apr 19;102(16):5820-5 PMID: 15811941
  10. Therapeutic gene silencing in neurological disorders, using interfering RNA.
    J Mol Med (Berl). 2005 Jun;83(6):413-9 PMID: 15759100
  11. Reversal of neuropathology and motor dysfunction in a conditional model of Huntington's disease.
    Cell. 2000 Mar 31;101(1):57-66 PMID: 10778856
  12. Decreased expression of striatal signaling genes in a mouse model of Huntington's disease.
    Hum Mol Genet. 2000 May 22;9(9):1259-71 PMID: 10814708
  13. The Huntington's disease protein interacts with p53 and CREB-binding protein and represses transcription.
    Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6763-8 PMID: 10823891
  14. Cannabinoid receptor messenger RNA levels decrease in a subset of neurons of the lateral striatum, cortex and hippocampus of transgenic Huntington's disease mice.
    Neuroscience. 2000;98(4):705-13 PMID: 10891614
  15. Transcriptional dysregulation in Huntington's disease.
    Trends Neurosci. 2000 Sep;23(9):387-92 PMID: 10941183
  16. Expanded polyglutamine stretches interact with TAFII130, interfering with CREB-dependent transcription.
    Nat Genet. 2000 Sep;26(1):29-36 PMID: 10973244
  17. Hammerhead ribozymes selectively suppress mutant type I collagen mRNA in osteogenesis imperfecta fibroblasts.
    Nucleic Acids Res. 2000 Oct 15;28(20):4013-20 PMID: 11024182
  18. Inactivation of Hdh in the brain and testis results in progressive neurodegeneration and sterility in mice.
    Nat Genet. 2000 Nov;26(3):300-6 PMID: 11062468
  19. Neurological abnormalities in a knock-in mouse model of Huntington's disease.
    Hum Mol Genet. 2001 Jan 15;10(2):137-44 PMID: 11152661
  20. Interaction of expanded polyglutamine stretches with nuclear transcription factors leads to aberrant transcriptional regulation in polyglutamine diseases.
    Neuropathology. 2000 Dec;20(4):326-33 PMID: 11211059
  21. Loss of normal huntingtin function: new developments in Huntington's disease research.
    Trends Neurosci. 2001 Mar;24(3):182-8 PMID: 11182459
  22. Protein kinase C beta II mRNA levels decrease in the striatum and cortex of transgenic Huntington's disease mice.
    J Psychiatry Neurosci. 2001 Mar;26(2):117-22 PMID: 11291528
  23. Human single-chain Fv intrabodies counteract in situ huntingtin aggregation in cellular models of Huntington's disease.
    Proc Natl Acad Sci U S A. 2001 Apr 10;98(8):4764-9 PMID: 11296304
  24. Loss of huntingtin-mediated BDNF gene transcription in Huntington's disease.
    Science. 2001 Jul 20;293(5529):493-8 PMID: 11408619
  25. Mice transgenic for exon 1 of the Huntington's disease gene display reduced striatal sensitivity to neurotoxicity induced by dopamine and 6-hydroxydopamine.
    Eur J Neurosci. 2001 Nov;14(9):1425-35 PMID: 11722604
  26. Resistance to NMDA toxicity correlates with appearance of nuclear inclusions, behavioural deficits and changes in calcium homeostasis in mice transgenic for exon 1 of the huntington gene.
    Eur J Neurosci. 2001 Nov;14(9):1492-504 PMID: 11722611
  27. Dysregulation of gene expression in the R6/2 model of polyglutamine disease: parallel changes in muscle and brain.
    Hum Mol Genet. 2002 Aug 15;11(17):1911-26 PMID: 12165554
  28. Early motor dysfunction and striosomal distribution of huntingtin microaggregates in Huntington's disease knock-in mice.
    J Neurosci. 2002 Sep 15;22(18):8266-76 PMID: 12223581
  29. Production and purification of serotype 1, 2, and 5 recombinant adeno-associated viral vectors.
    Methods. 2002 Oct;28(2):158-67 PMID: 12413414
  30. Evidence for dysfunction of the nigrostriatal pathway in the R6/1 line of transgenic Huntington's disease mice.
    Neurobiol Dis. 2002 Oct;11(1):134-46 PMID: 12460553
  31. Adeno-associated virus-mediated aspartoacylase gene transfer to the brain of knockout mouse for canavan disease.
    Mol Ther. 2003 May;7(5 Pt 1):580-7 PMID: 12718900
  32. Huntingtin aggregation and toxicity in Huntington's disease.
    Lancet. 2003 May 10;361(9369):1642-4 PMID: 12747895
  33. Allele-specific silencing of dominant disease genes.
    Proc Natl Acad Sci U S A. 2003 Jun 10;100(12):7195-200 PMID: 12782788
  34. Molecular and behavioral analysis of the R6/1 Huntington's disease transgenic mouse.
    Neuroscience. 2003;122(4):1049-57 PMID: 14643771
  35. Striatal phosphodiesterase mRNA and protein levels are reduced in Huntington's disease transgenic mice prior to the onset of motor symptoms.
    Neuroscience. 2004;123(4):967-81 PMID: 14751289
  36. Decreased cAMP response element-mediated transcription: an early event in exon 1 and full-length cell models of Huntington's disease that contributes to polyglutamine pathogenesis.
    J Biol Chem. 2004 Feb 6;279(6):4988-99 PMID: 14627700
  37. Molecular medicine for the brain: silencing of disease genes with RNA interference.
    Lancet Neurol. 2004 Mar;3(3):145-9 PMID: 14980529
  38. Huntingtin-protein interactions and the pathogenesis of Huntington's disease.
    Trends Genet. 2004 Mar;20(3):146-54 PMID: 15036808
  39. Hammerhead ribozyme targeting connective tissue growth factor mRNA blocks transforming growth factor-beta mediated cell proliferation.
    Exp Eye Res. 2004 Jun;78(6):1127-36 PMID: 15109919
  40. Inhibition of polyglutamine aggregate cytotoxicity by a structure-based elongation inhibitor.
    FASEB J. 2004 May;18(7):923-5 PMID: 15001566
  41. Huntington's disease: new paths to pathogenesis.
    Cell. 2004 Jul 9;118(1):4-7 PMID: 15242639
  42. Huntingtin controls neurotrophic support and survival of neurons by enhancing BDNF vesicular transport along microtubules.
    Cell. 2004 Jul 9;118(1):127-38 PMID: 15242649
  43. RNAi suppresses polyglutamine-induced neurodegeneration in a model of spinocerebellar ataxia.
    Nat Med. 2004 Aug;10(8):816-20 PMID: 15235598
  44. Recombinant AAV viral vectors pseudotyped with viral capsids from serotypes 1, 2, and 5 display differential efficiency and cell tropism after delivery to different regions of the central nervous system.
    Mol Ther. 2004 Aug;10(2):302-17 PMID: 15294177
  45. RNAi of COL1A1 in mesenchymal progenitor cells.
    Eur J Hum Genet. 2004 Oct;12(10):864-6 PMID: 15241481
  46. Inclusion body formation reduces levels of mutant huntingtin and the risk of neuronal death.
    Nature. 2004 Oct 14;431(7010):805-10 PMID: 15483602
  47. The new stereological tools: disector, fractionator, nucleator and point sampled intercepts and their use in pathological research and diagnosis.
    APMIS. 1988 Oct;96(10):857-81 PMID: 3056461
  48. A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington's disease chromosomes. The Huntington's Disease Collaborative Research Group.
    Cell. 1993 Mar 26;72(6):971-83 PMID: 8458085
  49. Polymorphism analysis of the huntingtin gene in Italian families affected with Huntington disease.
    Hum Mol Genet. 1994 Jul;3(7):1129-32 PMID: 7981682
  50. Widespread expression of Huntington's disease gene (IT15) protein product.
    Neuron. 1995 May;14(5):1065-74 PMID: 7748554
  51. Haplotype analysis of the delta 2642 and (CAG)n polymorphisms in the Huntington's disease (HD) gene provides an explanation for an apparent 'founder' HD haplotype.
    Hum Mol Genet. 1995 Feb;4(2):203-6 PMID: 7757068
  52. Inactivation of the mouse Huntington's disease gene homolog Hdh.
    Science. 1995 Jul 21;269(5222):407-10 PMID: 7618107
  53. Cellular localization of the Huntington's disease protein and discrimination of the normal and mutated form.
    Nat Genet. 1995 May;10(1):104-10 PMID: 7647777
  54. Increased apoptosis and early embryonic lethality in mice nullizygous for the Huntington's disease gene homologue.
    Nat Genet. 1995 Oct;11(2):155-63 PMID: 7550343
  55. Exon 1 of the HD gene with an expanded CAG repeat is sufficient to cause a progressive neurological phenotype in transgenic mice.
    Cell. 1996 Nov 1;87(3):493-506 PMID: 8898202
  56. Relationships of the 2642 deletion polymorphism (delta 2642) in the huntingtin gene with the CAG repeat expansion length and age at onset of the disease.
    Genet Couns. 1996;7(4):297-302 PMID: 8985734
  57. Instability of highly expanded CAG repeats in mice transgenic for the Huntington's disease mutation.
    Nat Genet. 1997 Feb;15(2):197-200 PMID: 9020849
  58. Formation of neuronal intranuclear inclusions underlies the neurological dysfunction in mice transgenic for the HD mutation.
    Cell. 1997 Aug 8;90(3):537-48 PMID: 9267033
  59. Huntingtin is required for neurogenesis and is not impaired by the Huntington's disease CAG expansion.
    Nat Genet. 1997 Dec;17(4):404-10 PMID: 9398841
  60. Characterization of intrastriatal recombinant adeno-associated virus-mediated gene transfer of human tyrosine hydroxylase and human GTP-cyclohydrolase I in a rat model of Parkinson's disease.
    J Neurosci. 1998 Jun 1;18(11):4271-84 PMID: 9592104
  61. Altered brain neurotransmitter receptors in transgenic mice expressing a portion of an abnormal human huntington disease gene.
    Proc Natl Acad Sci U S A. 1998 May 26;95(11):6480-5 PMID: 9600992
  62. Selective and rapid uptake of adeno-associated virus type 2 in brain.
    Hum Gene Ther. 1998 May 20;9(8):1181-6 PMID: 9625257
  63. Huntingtin acts in the nucleus to induce apoptosis but death does not correlate with the formation of intranuclear inclusions.
    Cell. 1998 Oct 2;95(1):55-66 PMID: 9778247
  64. Novel tools for production and purification of recombinant adenoassociated virus vectors.
    Hum Gene Ther. 1998 Dec 10;9(18):2745-60 PMID: 9874273
  65. Cloning and characterization of adeno-associated virus type 5.
    J Virol. 1999 Feb;73(2):1309-19 PMID: 9882336
  66. Intranuclear inclusions and neuritic aggregates in transgenic mice expressing a mutant N-terminal fragment of huntingtin.
    Hum Mol Genet. 1999 Mar;8(3):397-407 PMID: 9949199
  67. Viral gene delivery selectively restores feeding and prevents lethality of dopamine-deficient mice.
    Neuron. 1999 Jan;22(1):167-78 PMID: 10027299
  68. Characterization of progressive motor deficits in mice transgenic for the human Huntington's disease mutation.
    J Neurosci. 1999 Apr 15;19(8):3248-57 PMID: 10191337
  69. Recent advances on the pathogenesis of Huntington's disease.
    Exp Neurol. 1999 May;157(1):1-18 PMID: 10222105
Article Info
Journal
Molecular therapy : the journal of the American Society of Gene Therapy
Abbr.
Mol Ther
ISSN
1525-0016
DOI
10.1016/j.ymthe.2005.05.006
Published
2005-10-00
Pages
618-33
Language
English
Region
United States
NLM ID
100890581
PMCID
PMC2656966
Subset
IM
Grants
NINDS NIH HHS · F31 NS011182 · United States
NINDS NIH HHS · R01 NS048588 · United States
NINDS NIH HHS · R01 NS048588-02 · United States
NINDS NIH HHS · 5 F31 NS 011182-05 · United States
External Links
PubMed source DOI: 10.1016/j.ymthe.2005.05.006 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2