title: Comparison of hepatic transcription profiles of locked ribonucleic acid antisense oligonucleotides: evidence of distinct pathways contributing to non-target mediated toxicity in mice.
abstract: Development of LNA gapmers, antisense oligonucleotides used for efficient inhibition of target RNA expression, is limited by non-target mediated hepatotoxicity issues. In the present study, we investigated hepatic transcription profiles of mice administered non-toxic and toxic LNA gapmers. After repeated administration, a toxic LNA gapmer (TS-2), but not a non-toxic LNA gapmer (NTS-1), caused hepatocyte necrosis and increased serum alanine aminotransferase levels. Microarray data revealed that, in addition to gene expression patterns consistent with hepatotoxicity, 17 genes in the clathrin-mediated endocytosis (CME) pathway were altered in the TS-2 group. TS-2 significantly down-regulated myosin 1E (Myo1E), which is involved in release of clathrin-coated pits from plasma membranes. To map the earliest transcription changes associated with LNA gapmer-induced hepatotoxicity, a second microarray analysis was performed using NTS-1, TS-2, and a severely toxic LNA gapmer (HTS-3) at 8, 16, and 72 h following a single administration in mice. The only histopathological change observed was minor hepatic hypertrophy in all LNA groups across time points. NTS-1, but not 2 toxic LNA gapmers, increased immune response genes at 8 and 16 h but not at 72 h. TS-2 significantly perturbed the CME pathway only at 72 h, while Myo1E levels were decreased at all time points. In contrast, HTS-3 modulated DNA damage pathway genes at 8 and 16 h and also modulated the CME pathway genes (but not Myo1E) at 16 h. Our results may suggest that different LNAs modulate distinct transcriptional genes and pathways contributing to non-target mediated hepatotoxicity in mice.
authors: * Drug Safety Research and Development, Pfizer Inc., Groton, Connecticut 06340;
authors: Satoko
authors: S
authors: Kakiuchi-Kiyota
authors: Petra H
authors: PH
authors: Koza-Taylor
authors: Srinivasa R
authors: SR
authors: Mantena
authors: Linda F
authors: LF
authors: Nelms
authors: Ahmed E
authors: AE
authors: Enayetallah
authors: Brett D
authors: BD
authors: Hollingshead
authors: Andrew D
authors: AD
authors: Burdick
authors: Lori A
authors: LA
authors: Reed
authors: James A
authors: JA
authors: Warneke
authors: Lawrence O
authors: LO
authors: Whiteley
authors: Anne M
authors: AM
authors: Ryan
authors: Nagappan
authors: N
authors: Mathialagan
mesh_terms: D000818:Animals; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology / Q000235:genetics / Q000473:pathology*; D002966:Clathrin / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects* / Q000235:genetics; D020869:Gene Expression Profiling; D007279:Injections, Subcutaneous; D008099:Liver / Q000187:drug effects* / Q000473:pathology; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000633:toxicity*; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000633:toxicity*; D059467:Transcriptome / Q000187:drug effects*
chemical_list: D002966:Clathrin; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; C477371:locked nucleic acid
keywords: LNA gapmers; antisense oligonucleotide; clathrin-mediated endocytosis; hepatotoxicity; microarray analysis