%0 Journal Article
%A Miller, James R C
%A Pfister, Edith L
%A Liu, Wanzhao
%A Andre, Ralph
%A Träger, Ulrike
%A Kennington, Lori A
%A Lo, Kimberly
%A Dijkstra, Sipke
%A Macdonald, Douglas
%A Ostroff, Gary
%A Aronin, Neil
%A Tabrizi, Sarah J
%T Allele-Selective Suppression of Mutant Huntingtin in Primary Human Blood Cells.
%J Scientific reports
%V 7
%@ 2045-2322
%C London
%I Nature Publishing Group
%M DKFZ-2017-01078
%P 46740 -
%D 2017
%X Post-transcriptional gene silencing is a promising therapy for the monogenic, autosomal dominant, Huntington's disease (HD). However, wild-type huntingtin (HTT) has important cellular functions, so the ideal strategy would selectively lower mutant HTT while sparing wild-type. HD patients were genotyped for heterozygosity at three SNP sites, before phasing each SNP allele to wild-type or mutant HTT. Primary ex vivo myeloid cells were isolated from heterozygous patients and transfected with SNP-targeted siRNA, using glucan particles taken up by phagocytosis. Highly selective mRNA knockdown was achieved when targeting each allele of rs362331 in exon 50 of the HTT transcript; this selectivity was also present on protein studies. However, similar selectivity was not observed when targeting rs362273 or rs362307. Furthermore, HD myeloid cells are hyper-reactive compared to control. Allele-selective suppression of either wild-type or mutant HTT produced a significant, equivalent reduction in the cytokine response of HD myeloid cells to LPS, suggesting that wild-type HTT has a novel immune function. We demonstrate a sequential therapeutic process comprising genotyping and mutant HTT-linkage of SNPs, followed by personalised allele-selective suppression in a small patient cohort. We further show that allele-selectivity in ex vivo patient cells is highly SNP-dependent, with implications for clinical trial target selection.
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:28436437
%2 pmc:PMC5402279
%R 10.1038/srep46740
%U https://inrepo02.dkfz.de/record/120651