Antagonism of microRNA-122 in mice by systemically administered LNA-antimiR leads to up-regulation of a large set of predicted target mRNAs in the liver

TY - JOUR

T1 - Antagonism of microRNA-122 in mice by systemically administered LNA-antimiR leads to up-regulation of a large set of predicted target mRNAs in the liver

AU - Elmen, Joachim

AU - Lindow, Morten

AU - Silahtaroglu, Asli

AU - Bak, Mads

AU - Christensen, Mette

AU - Lind-Thomsen, Allan

AU - Hedtjarn, Maj

AU - hansen, jens bo

AU - Hansen, Henrik Freydenlund

AU - Straarup, Ellen Marie

AU - McCullagh, Keith

AU - Keraney, Phil

AU - Kauppinen, Markus Sakari

N1 - Paper id:: PMID: 18158304

PY - 2008

Y1 - 2008

N2 - MicroRNA-122 (miR-122) is an abundant liver-specific miRNA, implicated in fatty acid and cholesterol metabolism as well as hepatitis C viral replication. Here, we report that a systemically administered 16-nt, unconjugated LNA (locked nucleic acid)- antimiR oligonucleotide complementary to the 5’end of miR-122 leads to specific, dose-dependentsilencing of miR-122 and shows no hepatotoxicity in mice. Antagonism of miR-122 is due to formation of stable heteroduplexes between the LNA-antimiR and miR-122 as detected by northern analysis. Fluorescence in situ hybridization demonstrated uptake of the LNA-antimiR in mouse liver cells, which was accompanied by markedly reduced hybridization signals for mature miR-122 in treated mice. Functional antagonism of miR-122 was inferred from a low cholesterol phenotype and derepression within 24 h of 199 liver mRNAs showing significant enrichment for miR-122 seed matches in their 3’ UTRs. Expression profiling extended to 3 weeks after the last LNA-antimiR dose revealed that most of the changes in liver gene expression were normalized to saline control levels coinciding withnormalized miR-122 and plasma cholesterol levels. Combined, these data suggest that miRNA antagonists comprised of LNA are valuable tools for identifying miRNA targets in vivo and for studying the biological role of miRNAs and miRNAassociated gene-regulatory networks in a physiological context.

AB - MicroRNA-122 (miR-122) is an abundant liver-specific miRNA, implicated in fatty acid and cholesterol metabolism as well as hepatitis C viral replication. Here, we report that a systemically administered 16-nt, unconjugated LNA (locked nucleic acid)- antimiR oligonucleotide complementary to the 5’end of miR-122 leads to specific, dose-dependentsilencing of miR-122 and shows no hepatotoxicity in mice. Antagonism of miR-122 is due to formation of stable heteroduplexes between the LNA-antimiR and miR-122 as detected by northern analysis. Fluorescence in situ hybridization demonstrated uptake of the LNA-antimiR in mouse liver cells, which was accompanied by markedly reduced hybridization signals for mature miR-122 in treated mice. Functional antagonism of miR-122 was inferred from a low cholesterol phenotype and derepression within 24 h of 199 liver mRNAs showing significant enrichment for miR-122 seed matches in their 3’ UTRs. Expression profiling extended to 3 weeks after the last LNA-antimiR dose revealed that most of the changes in liver gene expression were normalized to saline control levels coinciding withnormalized miR-122 and plasma cholesterol levels. Combined, these data suggest that miRNA antagonists comprised of LNA are valuable tools for identifying miRNA targets in vivo and for studying the biological role of miRNAs and miRNAassociated gene-regulatory networks in a physiological context.

U2 - 10.1093/nar/gkm1113

DO - 10.1093/nar/gkm1113

M3 - Journal article

SN - 0305-1048

VL - 36

SP - 1153

EP - 1162

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 4

ER -