Potent and selective antisense oligonucleotides targeting single-nucleotide polymorphisms in the Huntington disease gene / allele-specific silencing of mutant huntingtin

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

SN - 1525-0016

VL - 19

SP - 2178

EP - 2185

JO - Molecular Therapy

JF - Molecular Therapy

IS - 12

ER -