title: Rational design of antisense oligonucleotides targeting single nucleotide polymorphisms for potent and allele selective suppression of mutant Huntingtin in the CNS.
abstract: Autosomal dominant diseases such as Huntington's disease (HD) are caused by a gain of function mutant protein and/or RNA. An ideal treatment for these diseases is to selectively suppress expression of the mutant allele while preserving expression of the wild-type variant. RNase H active antisense oligonucleotides (ASOs) or small interfering RNAs can achieve allele selective suppression of gene expression by targeting single nucleotide polymorphisms (SNPs) associated with the repeat expansion. ASOs have been previously shown to discriminate single nucleotide changes in targeted RNAs with ∼5-fold selectivity. Based on RNase H enzymology, we enhanced single nucleotide discrimination by positional incorporation of chemical modifications within the oligonucleotide to limit RNase H cleavage of the non-targeted transcript. The resulting oligonucleotides demonstrate >100-fold discrimination for a single nucleotide change at an SNP site in the disease causing huntingtin mRNA, in patient cells and in a completely humanized mouse model of HD. The modified ASOs were also well tolerated after injection into the central nervous system of wild-type animals, suggesting that their tolerability profile is suitable for advancement as potential allele-selective HD therapeutics. Our findings lay the foundation for efficient allele-selective downregulation of gene expression using ASOs-an outcome with broad application to HD and other dominant genetic disorders.
authors: Isis Pharmaceuticals, 2855 Gazelle Court, Carlsbad, CA 92010, USA and Centre for Molecular Medicine and Therapeutics, Child and Family Research Institute, University of British Columbia, Vancouver, British Columbia, V5Z 4H4, Canada.
authors: Michael E
authors: ME
authors: Østergaard
authors: Amber L
authors: AL
authors: Southwell
authors: Holly
authors: H
authors: Kordasiewicz
authors: Andrew T
authors: AT
authors: Watt
authors: Niels H
authors: NH
authors: Skotte
authors: Crystal N
authors: CN
authors: Doty
authors: Kuljeet
authors: K
authors: Vaid
authors: Erika B
authors: EB
authors: Villanueva
authors: Eric E
authors: EE
authors: Swayze
authors: C Frank
authors: CF
authors: Bennett
authors: Michael R
authors: MR
authors: Hayden
authors: Punit P
authors: PP
authors: Seth
mesh_terms: D000483:Alleles; D000818:Animals; D020029:Base Pairing; D001921:Brain / Q000378:metabolism; D002478:Cells, Cultured; D015536:Down-Regulation; D005461:Fluorine / Q000737:chemistry; D006801:Humans; D000071058:Huntingtin Protein; D006816:Huntington Disease / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000737:chemistry*; D020641:Polymorphism, Single Nucleotide; D051381:Rats; D017207:Rats, Sprague-Dawley; D016914:Ribonuclease H / Q000378:metabolism
chemical_list: C086055:HTT protein, human; D000071058:Huntingtin Protein; D009419:Nerve Tissue Proteins; D016376:Oligonucleotides, Antisense; D005461:Fluorine; D016914:Ribonuclease H; C078856:ribonuclease HI
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