Unexpected off-targeting effects of anti-huntingtin ribozymes and siRNA in vivo
Gene transfer strategies to reduce levels of mutant huntingtin (mHtt) mRNA and protein by targeting human Htt have shown therapeutic promise in vivo. Previously, we have reported that a specific, adeno-associated viral vector (rAAV)-delivered short-hairpin RNA (siHUNT-2) targeting human Htt mRNA une...
Main Authors: | , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Elsevier
2008-03-01
|
Series: | Neurobiology of Disease |
Subjects: | |
Online Access: | http://www.sciencedirect.com/science/article/pii/S0969996107002495 |
id |
doaj-b9ff9b59dbae4af3b27be35340e8eb41 |
---|---|
record_format |
Article |
spelling |
doaj-b9ff9b59dbae4af3b27be35340e8eb412021-03-20T04:55:15ZengElsevierNeurobiology of Disease1095-953X2008-03-01293446455Unexpected off-targeting effects of anti-huntingtin ribozymes and siRNA in vivoEileen M. Denovan-Wright0Edgardo Rodriguez-Lebron1Alfred S. Lewin2Ronald J. Mandel3Department of Pharmacology, Dalhousie University, Halifax, Nova Scotia, Canada B3H 1X5Department of Neurology, University of Michigan, Ann Arbor, MI, USADepartment of Molecular Genetics and Microbiology, University of Florida, College of Medicine, Gainesville, FL, USADepartment of Neuroscience, University of Florida McKnight Brain Institute and Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, FL, USA; Corresponding author. Fax: +1 352 392 8347.Gene transfer strategies to reduce levels of mutant huntingtin (mHtt) mRNA and protein by targeting human Htt have shown therapeutic promise in vivo. Previously, we have reported that a specific, adeno-associated viral vector (rAAV)-delivered short-hairpin RNA (siHUNT-2) targeting human Htt mRNA unexpectedly decreased levels of striatal-specific transcripts in both wild-type and R6/1 transgenic HD mice. The goal of this study was to determine whether the siHUNT-2-mediated effect was due to adverse effects of RNA interference (RNAi) expression in the brain. To this end, we designed two catalytically active hammerhead ribozymes directed against the same region of human Htt mRNA targeted by siHUNT-2 and delivered them to wild-type and R6/1 transgenic HD mice. After 10 weeks of continuous expression, these ribozymes, like siHUNT-2, negatively impacted the expression of a subset of genes in the striatum. This effect was independent of rAAV transduction and specific to the targeting of a unique sequence in human Htt mRNA. After consideration of the known potential RNAi-specific toxic mechanisms, only cleavage of an unintended RNA target can account for the data reported herein. Thus, long-term rAAV-mediated RNAi in the brain does not, in and of itself, negatively affect striatal gene expression. These findings have important implications in the development of therapeutic RNAi for the treatment of neurological disease.http://www.sciencedirect.com/science/article/pii/S0969996107002495rAAVStriatumHuntington’s diseaseMouseGene transferNeurological disorder |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Eileen M. Denovan-Wright Edgardo Rodriguez-Lebron Alfred S. Lewin Ronald J. Mandel |
spellingShingle |
Eileen M. Denovan-Wright Edgardo Rodriguez-Lebron Alfred S. Lewin Ronald J. Mandel Unexpected off-targeting effects of anti-huntingtin ribozymes and siRNA in vivo Neurobiology of Disease rAAV Striatum Huntington’s disease Mouse Gene transfer Neurological disorder |
author_facet |
Eileen M. Denovan-Wright Edgardo Rodriguez-Lebron Alfred S. Lewin Ronald J. Mandel |
author_sort |
Eileen M. Denovan-Wright |
title |
Unexpected off-targeting effects of anti-huntingtin ribozymes and siRNA in vivo |
title_short |
Unexpected off-targeting effects of anti-huntingtin ribozymes and siRNA in vivo |
title_full |
Unexpected off-targeting effects of anti-huntingtin ribozymes and siRNA in vivo |
title_fullStr |
Unexpected off-targeting effects of anti-huntingtin ribozymes and siRNA in vivo |
title_full_unstemmed |
Unexpected off-targeting effects of anti-huntingtin ribozymes and siRNA in vivo |
title_sort |
unexpected off-targeting effects of anti-huntingtin ribozymes and sirna in vivo |
publisher |
Elsevier |
series |
Neurobiology of Disease |
issn |
1095-953X |
publishDate |
2008-03-01 |
description |
Gene transfer strategies to reduce levels of mutant huntingtin (mHtt) mRNA and protein by targeting human Htt have shown therapeutic promise in vivo. Previously, we have reported that a specific, adeno-associated viral vector (rAAV)-delivered short-hairpin RNA (siHUNT-2) targeting human Htt mRNA unexpectedly decreased levels of striatal-specific transcripts in both wild-type and R6/1 transgenic HD mice. The goal of this study was to determine whether the siHUNT-2-mediated effect was due to adverse effects of RNA interference (RNAi) expression in the brain. To this end, we designed two catalytically active hammerhead ribozymes directed against the same region of human Htt mRNA targeted by siHUNT-2 and delivered them to wild-type and R6/1 transgenic HD mice. After 10 weeks of continuous expression, these ribozymes, like siHUNT-2, negatively impacted the expression of a subset of genes in the striatum. This effect was independent of rAAV transduction and specific to the targeting of a unique sequence in human Htt mRNA. After consideration of the known potential RNAi-specific toxic mechanisms, only cleavage of an unintended RNA target can account for the data reported herein. Thus, long-term rAAV-mediated RNAi in the brain does not, in and of itself, negatively affect striatal gene expression. These findings have important implications in the development of therapeutic RNAi for the treatment of neurological disease. |
topic |
rAAV Striatum Huntington’s disease Mouse Gene transfer Neurological disorder |
url |
http://www.sciencedirect.com/science/article/pii/S0969996107002495 |
work_keys_str_mv |
AT eileenmdenovanwright unexpectedofftargetingeffectsofantihuntingtinribozymesandsirnainvivo AT edgardorodriguezlebron unexpectedofftargetingeffectsofantihuntingtinribozymesandsirnainvivo AT alfredslewin unexpectedofftargetingeffectsofantihuntingtinribozymesandsirnainvivo AT ronaldjmandel unexpectedofftargetingeffectsofantihuntingtinribozymesandsirnainvivo |
_version_ |
1724211785097019392 |