Allele-Selective Inhibition of Mutant Huntingtin Expression with Antisense Oligonucleotides Targeting the Expanded CAG Repeat

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• 作者：Keith T. Gagnon ; Hannah M. Pendergraff ; Glen F. Deleavey ; Eric E. Swayze ; Pierre Potier ; John Randolph ; Eric B. Roesch ; Jyoti Chattopadhyaya ; Masad J. Damha ; C. Frank Bennett ; Christophe Montaillier
• 刊名：Biochemistry
• 出版年：2010
• 出版时间：November 30, 2010
• 年：2010
• 卷：49
• 期：47
• 页码：10166-10178
• 全文大小：1274K
• 年卷期：v.49,no.47(November 30, 2010)
• ISSN：1520-4995

文摘

Huntington’s disease (HD) is a currently incurable neurodegenerative disease caused by the expansion of a CAG trinucleotide repeat within the huntingtin (HTT) gene. Therapeutic approaches include selectively inhibiting the expression of the mutated HTT allele while conserving function of the normal allele. We have evaluated a series of antisense oligonucleotides (ASOs) targeted to the expanded CAG repeat within HTT mRNA for their ability to selectively inhibit expression of mutant HTT protein. Several ASOs incorporating a variety of modifications, including bridged nucleic acids and phosphorothioate internucleotide linkages, exhibited allele-selective silencing in patient-derived fibroblasts. Allele-selective ASOs did not affect the expression of other CAG repeat-containing genes and selectivity was observed in cell lines containing minimal CAG repeat lengths representative of most HD patients. Allele-selective ASOs left HTT mRNA intact and did not support ribonuclease H activity in vitro. We observed cooperative binding of multiple ASO molecules to CAG repeat-containing HTT mRNA transcripts in vitro. These results are consistent with a mechanism involving inhibition at the level of translation. ASOs targeted to the CAG repeat of HTT provide a starting point for the development of oligonucleotide-based therapeutics that can inhibit gene expression with allelic discrimination in patients with HD.