Journal Article

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

Chandhok, Gursimran, Sauer, Vanessa, Stella, Jacqueline, Guttmann, Sarah, Guo, Shuling, Gollob, Jared, Monia, Brett P., Zibert, Andree, Avsar, Yesim, Fleischhauer, Lutz, Niemietz, Christoph J., Ackermann, Elizabeth J., Schmidt, Hartmut H. -J.

2016

Overview

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.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Adult / Aged / Amyloid / Amyloid Neuropathies, Familial - drug therapy / Amyloid Neuropathies, Familial - genetics / Amyloid Neuropathies, Familial - urine / Amyloidosis / Animal models / Antisense oligonucleotides / Biology and Life Sciences / Cardiomyopathy / Care and treatment / Cell culture / Cell cycle / Cell Differentiation / Clinical trials / Disease / Drug Evaluation, Preclinical / Female / Fibroblasts / Gene expression / Gene Knockdown Techniques / Health aspects / Hepatocytes / Hepatocytes - drug effects / Humans / Induced Pluripotent Stem Cells - cytology / Liver / Male / Medical research / Medicine and Health Sciences / Middle Aged / Mutation / Oligonucleotides / Oligonucleotides, Antisense - pharmacology / Oligonucleotides, Antisense - therapeutic use / Patients / Pharmaceuticals / Pluripotency / Polyneuropathy / Prealbumin - genetics / Research and Analysis Methods / Ribonucleic acid / RNA / RNA, Messenger - genetics / RNA, Small Interfering - genetics / siRNA / Stem cell transplantation / Stem cells / Tissue culture / Transthyretin / Urine