Potent and selective antisense oligonucleotides targeting single-nucleotide polymorphisms in the Huntington disease gene / allele-specific silencing of mutant huntingtin
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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Potent and selective antisense oligonucleotides targeting single-nucleotide polymorphisms in the Huntington disease gene / allele-specific silencing of mutant huntingtin. / Carroll, Jeffrey B; Warby, Simon C; Southwell, Amber L; Doty, Crystal N; Greenlee, Sarah; Skotte, Niels; Hung, Gene; Bennett, C Frank; Freier, Susan M; Hayden, Michael R.
I: Molecular therapy : the journal of the American Society of Gene Therapy, Bind 19, Nr. 12, 12.2011, s. 2178-85.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Potent and selective antisense oligonucleotides targeting single-nucleotide polymorphisms in the Huntington disease gene / allele-specific silencing of mutant huntingtin
AU - Carroll, Jeffrey B
AU - Warby, Simon C
AU - Southwell, Amber L
AU - Doty, Crystal N
AU - Greenlee, Sarah
AU - Skotte, Niels
AU - Hung, Gene
AU - Bennett, C Frank
AU - Freier, Susan M
AU - Hayden, Michael R
PY - 2011/12
Y1 - 2011/12
N2 - Huntington disease (HD) is an autosomal dominant neurodegenerative disorder caused by CAG-expansion in the huntingtin gene (HTT) that results in a toxic gain of function in the mutant huntingtin protein (mHTT). Reducing the expression of mHTT is therefore an attractive therapy for HD. However, wild-type HTT protein is essential for development and has critical roles in maintaining neuronal health. Therapies for HD that reduce wild-type HTT may therefore generate unintended negative consequences. We have identified single-nucleotide polymorphism (SNP) targets in the human HD population for the disease-specific targeting of the HTT gene. Using primary cells from patients with HD and the transgenic YAC18 and BACHD mouse lines, we developed antisense oligonucleotide (ASO) molecules that potently and selectively silence mHTT at both exonic and intronic SNP sites. Modification of these ASOs with S-constrained-ethyl (cET) motifs significantly improves potency while maintaining allele selectively in vitro. The developed ASO is potent and selective for mHTT in vivo after delivery to the mouse brain. We demonstrate that potent and selective allele-specific knockdown of the mHTT protein can be achieved at therapeutically relevant SNP sites using ASOs in vitro and in vivo.
AB - Huntington disease (HD) is an autosomal dominant neurodegenerative disorder caused by CAG-expansion in the huntingtin gene (HTT) that results in a toxic gain of function in the mutant huntingtin protein (mHTT). Reducing the expression of mHTT is therefore an attractive therapy for HD. However, wild-type HTT protein is essential for development and has critical roles in maintaining neuronal health. Therapies for HD that reduce wild-type HTT may therefore generate unintended negative consequences. We have identified single-nucleotide polymorphism (SNP) targets in the human HD population for the disease-specific targeting of the HTT gene. Using primary cells from patients with HD and the transgenic YAC18 and BACHD mouse lines, we developed antisense oligonucleotide (ASO) molecules that potently and selectively silence mHTT at both exonic and intronic SNP sites. Modification of these ASOs with S-constrained-ethyl (cET) motifs significantly improves potency while maintaining allele selectively in vitro. The developed ASO is potent and selective for mHTT in vivo after delivery to the mouse brain. We demonstrate that potent and selective allele-specific knockdown of the mHTT protein can be achieved at therapeutically relevant SNP sites using ASOs in vitro and in vivo.
KW - Alleles
KW - Animals
KW - Brain
KW - Cells, Cultured
KW - Female
KW - Fibroblasts
KW - Gene Silencing
KW - Genetic Therapy
KW - Humans
KW - Huntingtin Protein
KW - Huntington Disease
KW - Male
KW - Mice
KW - Mice, Transgenic
KW - Mutant Proteins
KW - Nerve Tissue Proteins
KW - Neurons
KW - Nuclear Proteins
KW - Oligonucleotides, Antisense
KW - Pedigree
KW - Polymorphism, Single Nucleotide
KW - RNA, Messenger
KW - Serotonin Plasma Membrane Transport Proteins
KW - Trinucleotide Repeat Expansion
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1038/mt.2011.201
DO - 10.1038/mt.2011.201
M3 - Journal article
C2 - 21971427
VL - 19
SP - 2178
EP - 2185
JO - Molecular Therapy
JF - Molecular Therapy
SN - 1525-0016
IS - 12
ER -
ID: 170597568