A majority of Huntington's disease patients may be treatable by individualized allele-specific RNA interference

• Published in: Experimental neurology Vol. 217; no. 2; pp. 312 - 319
• Main Authors: Lombardi, Maria Stella, Jaspers, Leonie, Spronkmans, Christine, Gellera, Cinzia, Taroni, Franco, Di Maria, Emilio, Donato, Stefano Di, Kaemmerer, William F.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Inc 01.06.2009; Elsevier
• Subjects: Adolescent; Adult; Aged; Aged, 80 and over; Allele-specific; Alleles; Biological and medical sciences; Child; Cohort Studies; Degenerative and inherited degenerative diseases of the nervous system. Leukodystrophies. Prion diseases; DNA Mutational Analysis; Female; Gene Frequency; Gene Targeting - methods; Genetic Testing; Genetic Therapy - methods; Haplotype; Heterozygote; Humans; Huntington Disease - genetics; Huntington Disease - physiopathology; Huntington Disease - therapy; Huntington's disease; Male; Medical sciences; Middle Aged; Neurology; Polymorphism, Single Nucleotide - genetics; Pyrosequencing; RNA interference; RNA Interference - physiology; RNA, Small Interfering - therapeutic use; Small interfering RNA; Young Adult; Small interfering RNA; RNA interference; Huntington's disease; Allele-specific; Haplotype; Pyrosequencing; Human; Nervous system diseases; Huntington disease; Interference; Genetic disease; Cerebral disorder; Central nervous system disease; Degenerative disease; Extrapyramidal syndrome
• ISSN: 0014-4886; 1090-2430
• DOI: 10.1016/j.expneurol.2009.03.004

Use of RNA interference to reduce huntingtin protein (htt) expression in affected brain regions may provide an effective treatment for Huntington disease (HD), but it remains uncertain whether suppression of both wild-type and mutant alleles in a heterozygous patient will provide more benefit than harm. Previous research has shown suppression of just the mutant allele is achievable using siRNA targeted to regions of HD mRNA containing single nucleotide polymorphisms (SNPs). To determine whether more than a minority of patients may be eligible for an allele-specific therapy, we genotyped DNA from 327 unrelated European Caucasian HD patients at 26 SNP sites in the HD gene. Over 86% of the patients were found to be heterozygous for at least one SNP among those tested. Because the sites are genetically linked, one cannot use the heterozygosity rates of the individual SNPs to predict how many sites (and corresponding allele-specific siRNA) would be needed to provide at least one treatment possibility for this percentage of patients. By computing all combinations, we found that a repertoire of allele-specific siRNA corresponding to seven sites can provide at least one allele-specific siRNA treatment option for 85.6% of our sample. Moreover, we provide evidence that allele-specific siRNA targeting these sites are readily identifiable using a high throughput screening method, and that allele-specific siRNA identified using this method indeed show selective suppression of endogenous mutant htt protein in fibroblast cells from HD patients. Therefore, allele-specific siRNA are not so rare as to be impractical to find and use therapeutically.