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Evaluation of Therapeutic Oligonucleotides for Familial Amyloid Polyneuropathy in Patient-Derived Hepatocyte-Like Cells
- Source :
- PLoS ONE, Vol 11, Iss 9, p e0161455 (2016), PLoS ONE
- Publication Year :
- 2016
- Publisher :
- Public Library of Science (PLoS), 2016.
-
Abstract
- 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.
- Subjects :
- Male
Physiology
Oligonucleotides
Drug Evaluation, Preclinical
lcsh:Medicine
Urine
Biochemistry
Animal Cells
Medicine and Health Sciences
Prealbumin
Small interfering RNAs
RNA, Small Interfering
lcsh:Science
Nucleotides
Stem Cells
Cell Differentiation
Middle Aged
Body Fluids
Nucleic acids
Liver
Gene Knockdown Techniques
Medicine and health
Female
Biological Cultures
Cellular Types
Anatomy
Research Article
Adult
endocrine system
Induced Pluripotent Stem Cells
Research and Analysis Methods
Genetics
Humans
RNA, Messenger
ddc:610
Non-coding RNA
Aged
Amyloid Neuropathies, Familial
lcsh:R
Biology and Life Sciences
nutritional and metabolic diseases
Cell Biology
Cell Cultures
Oligonucleotides, Antisense
Gene regulation
Hepatocytes
RNA
lcsh:Q
Gene expression
Developmental Biology
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 11
- Issue :
- 9
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.pmid.dedup....069d1baf845a211e4af6e0e67f8525ad