title: Evaluation of Therapeutic Oligonucleotides for Familial Amyloid Polyneuropathy in Patient-Derived Hepatocyte-Like Cells.
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.
authors: Klinik für Transplantationsmedizin, Universitätsklinikum Münster, Albert-Schweitzer-Campus 1, Gebäude A14, D-48149, Münster, Germany.
authors: Christoph J
authors: CJ
authors: Niemietz
authors: Klinik für Transplantationsmedizin, Universitätsklinikum Münster, Albert-Schweitzer-Campus 1, Gebäude A14, D-48149, Münster, Germany.
authors: Vanessa
authors: V
authors: Sauer
authors: Klinik für Transplantationsmedizin, Universitätsklinikum Münster, Albert-Schweitzer-Campus 1, Gebäude A14, D-48149, Münster, Germany.
authors: Jacqueline
authors: J
authors: Stella
authors: Klinik für Transplantationsmedizin, Universitätsklinikum Münster, Albert-Schweitzer-Campus 1, Gebäude A14, D-48149, Münster, Germany.
authors: Lutz
authors: L
authors: Fleischhauer
authors: Klinik für Transplantationsmedizin, Universitätsklinikum Münster, Albert-Schweitzer-Campus 1, Gebäude A14, D-48149, Münster, Germany.
authors: Gursimran
authors: G
authors: Chandhok
authors: Klinik für Transplantationsmedizin, Universitätsklinikum Münster, Albert-Schweitzer-Campus 1, Gebäude A14, D-48149, Münster, Germany.
authors: Sarah
authors: S
authors: Guttmann
authors: Klinik für Transplantationsmedizin, Universitätsklinikum Münster, Albert-Schweitzer-Campus 1, Gebäude A14, D-48149, Münster, Germany.
authors: Yesim
authors: Y
authors: Avsar
authors: Ionis Pharmaceuticals, Inc., 2855 Gazelle Ct, Carlsbad, CA, 92010, United States of America.
authors: Shuling
authors: S
authors: Guo
authors: Ionis Pharmaceuticals, Inc., 2855 Gazelle Ct, Carlsbad, CA, 92010, United States of America.
authors: Elizabeth J
authors: EJ
authors: Ackermann
authors: Alnylam Pharmaceuticals, Inc., 300 Third St #3, Cambridge, MA, 02142, United States of America.
authors: Jared
authors: J
authors: Gollob
authors: Ionis Pharmaceuticals, Inc., 2855 Gazelle Ct, Carlsbad, CA, 92010, United States of America.
authors: Brett P
authors: BP
authors: Monia
authors: Klinik für Transplantationsmedizin, Universitätsklinikum Münster, Albert-Schweitzer-Campus 1, Gebäude A14, D-48149, Münster, Germany.
authors: Andree
authors: A
authors: Zibert
authors: Klinik für Transplantationsmedizin, Universitätsklinikum Münster, Albert-Schweitzer-Campus 1, Gebäude A14, D-48149, Münster, Germany.
authors: Hartmut H-J
authors: HH
authors: Schmidt
mesh_terms: D000328:Adult; D000368:Aged; D028227:Amyloid Neuropathies, Familial / Q000188:drug therapy* / Q000235:genetics / Q000652:urine; D002454:Cell Differentiation; D004353:Drug Evaluation, Preclinical; D005260:Female; D055785:Gene Knockdown Techniques; D022781:Hepatocytes / Q000187:drug effects*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology; D008297:Male; D008875:Middle Aged; D016376:Oligonucleotides, Antisense / Q000494:pharmacology / Q000627:therapeutic use*; D011228:Prealbumin / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics
chemical_list: D016376:Oligonucleotides, Antisense; D011228:Prealbumin; D012333:RNA, Messenger; D034741:RNA, Small Interfering