Abstract
Antisense oligonucleotides (ASO) containing 2′-O-methoxyethyl ribose (2′-MOE) modifications have been shown to possess both excellent pharmacokinetic properties and robust pharmacological activity in several animal models of human disease. 2′-MOE ASOs are generally well tolerated, displaying minimal to mild proinflammatory effect at doses far exceeding therapeutic doses. Although the vast majority of 2′-MOE ASOs are safe and well tolerated, a small subset of ASOs inducing acute inflammation in mice has been identified. The mechanism for these findings is not clear at this point, but the effects are clearly sequence-specific. One of those ASOs, ISIS 147420, causes a severe inflammatory response atypical of this class of oligonucleotides characterized by induction in interferon-β (IFN-β) at 48 h followed by acute transaminitis and extensive hepatocyte apoptosis and necrosis at 72 h. A large number of interferon-stimulated genes were significantly up-regulated in liver as early as 24 h. We speculated that a specific sequence motif might cause ISIS 147420 to be mistaken for viral RNA or DNA, thus triggering an acute innate immune response. ISIS 147420 toxicity was independent of Toll-like receptors, because there was no decrease in IFN-β in Toll/interleukin-1 receptor-domain-containing adapter-inducing IFN-β or Myd88-deficient mice. The involvement of cytosolic retinoic acid-inducible gene (RIG)-I-like pattern recognition receptors was also investigated. Pretreatment of mice with melanoma differentiation-associated gene 5 (MDA5) and IFN-β promoter stimulator-1 ASOs, but not RIG-I or laboratory of genetics and physiology 2 (LGP2) ASOs, prevented the increase in IFN-β and alanine aminotransferase induced by ISIS 147420. These results revealed a novel mechanism of oligonucleotide-mediated toxicity requiring both MDA5 and IPS-1 and resulting in the activation of the innate immune response.
Footnotes
Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
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The online version of this article (available at http://jpet.aspetjournals.org) contains supplemental material.
ABBREVIATIONS:
- ASO
- antisense oligonucleotide
- PS
- phosphorothioate
- MOE
- O-methoxyethyl ribose
- ALT
- alanine aminotransferase
- AST
- aspartate aminotransferase
- PRR
- pattern recognition receptor
- TLR
- Toll-like receptor
- FDR
- false discovery rate
- MDA5
- melanoma differentiation-associated gene 5
- RIG
- retinoic acid-inducible gene
- RLR
- RIG-I-like receptor
- IFN
- interferon
- IPS-1
- IFN-β promoter stimulator-1
- TRIF
- Toll/interleukin-1 receptor-domain-containing adapter-inducing IFN-β
- MYD88
- myeloid differentiation primary response gene 88
- KO
- knockout
- IPA
- Ingenuity Pathway Analysis
- TBST
- Tris-buffered saline/Tween 20
- ANOVA
- analysis of variance
- SDM
- standard deviation of mean
- STAT1
- signal transducer and activator of transcription 1
- HMGB
- high-mobility group box
- LGP2
- laboratory of genetics and physiology 2.
- Received March 5, 2012.
- Accepted April 12, 2012.
- Copyright © 2012 by The American Society for Pharmacology and Experimental Therapeutics
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