Evaluation of Therapeutic Oligonucleotides for Familial Amyloid Polyneuropathy in Patient-Derived Hepatocyte-Like Cells

Familial amyloid polyneuropathy (FAP) is caused by mutations of the transthyretin (TTR) gene, predominantly expressed in the liver. Two compounds that knockdown TTR, comprising a small interfering RNA (siRNA; ALN-TTR-02) and an antisense oligonucleotide (ASO; IONIS-TTRRx), are currently being evalua...

Authors: Niemietz, Christoph
Sauer, Vanessa Daniela
Stella, Jaqueline
Fleischhauer, Lutz
Chandhok, Gursimran
Guttmann, Sarah
Avsar, Yesim
Guo, Shuling
Ackermann, Elisabeth J.
Gollob, Jared
Monia, Brett P.
Zibert, Andree
Schmidt, Hartmut
Division/Institute:FB 05: Medizinische Fakultät
Document types:Article
Media types:Text
Publication date:2016
Date of publication on miami:18.01.2017
Modification date:23.01.2020
Edition statement:[Electronic ed.]
Source:PLoS ONE 11 (2016) 9, e0161455, 1-16
DDC Subject:610: Medizin und Gesundheit
License:CC BY 4.0
Language:English
Notes:Finanziert durch den Open-Access-Publikationsfonds 2015/2016 der Westfälischen Wilhelms-Universität Münster (WWU Münster)
Format:PDF document
ISSN:1932-6203
URN:urn:nbn:de:hbz:6-83249600713
Permalink:http://nbn-resolving.de/urn:nbn:de:hbz:6-83249600713
Other Identifiers:DOI: 10.1371/journal.pone.0161455
Digital documents:journal.pone.0161455.PDF

Familial amyloid polyneuropathy (FAP) is caused by mutations of the transthyretin (TTR) gene, predominantly expressed in the liver. Two compounds that knockdown TTR, comprising a small interfering RNA (siRNA; ALN-TTR-02) and an antisense oligonucleotide (ASO; IONIS-TTRRx), are currently being evaluated in clinical trials. Since primary hepatocytes from FAP patients are rarely available for molecular analysis and commercial tissue culture cells or animal models lack the patient-specific genetic background, this study uses primary cells derived from urine of FAP patients. Urine-derived cells were reprogrammed to induced pluripotent stem cells (iPSCs) with high efficiency. Hepatocyte-like cells (HLCs) showing typical hepatic marker expression were obtained from iPSCs of the FAP patients. TTR mRNA expression of FAP HLCs almost reached levels measured in human hepatocytes. To assess TTR knockdown, siTTR1 and TTR-ASO were introduced to HLCs. A significant downregulation (>80%) of TTR mRNA was induced in the HLCs by both oligonucleotides. TTR protein present in the cell culture supernatant of HLCs was similarly downregulated. Gene expression of other hepatic markers was not affected by the therapeutic oligonucleotides. Our data indicate that urine cells (UCs) after reprogramming and hepatic differentiation represent excellent primary human target cells to assess the efficacy and specificity of novel compounds.