TY  - JOUR
AU  - Miller, James R C
AU  - Pfister, Edith L
AU  - Liu, Wanzhao
AU  - Andre, Ralph
AU  - Träger, Ulrike
AU  - Kennington, Lori A
AU  - Lo, Kimberly
AU  - Dijkstra, Sipke
AU  - Macdonald, Douglas
AU  - Ostroff, Gary
AU  - Aronin, Neil
AU  - Tabrizi, Sarah J
TI  - Allele-Selective Suppression of Mutant Huntingtin in Primary Human Blood Cells.
JO  - Scientific reports
VL  - 7
SN  - 2045-2322
CY  - London
PB  - Nature Publishing Group
M1  - DKFZ-2017-01078
SP  - 46740 -
PY  - 2017
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - pmid:28436437
C2  - pmc:PMC5402279
DO  - DOI:10.1038/srep46740
UR  - https://inrepo02.dkfz.de/record/120651
ER  -