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.
MeSH terms
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Adaptor Proteins, Signal Transducing / genetics
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Adaptor Proteins, Signal Transducing / immunology
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Adaptor Proteins, Signal Transducing / metabolism
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Adaptor Proteins, Vesicular Transport / genetics
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Adaptor Proteins, Vesicular Transport / immunology
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Adaptor Proteins, Vesicular Transport / metabolism
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Alanine Transaminase / genetics
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Alanine Transaminase / immunology
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Alanine Transaminase / metabolism
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Animals
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DEAD-box RNA Helicases / genetics
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DEAD-box RNA Helicases / immunology*
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DEAD-box RNA Helicases / metabolism
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DNA / genetics
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DNA / immunology*
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DNA / metabolism
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Hepatocytes / immunology
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Hepatocytes / metabolism
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Immunity, Innate / genetics
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Immunity, Innate / immunology*
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Inflammation / genetics
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Inflammation / immunology
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Inflammation / metabolism
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Interferon Type I / genetics
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Interferon Type I / immunology*
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Interferon Type I / metabolism
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Interferon-Induced Helicase, IFIH1
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Interferon-beta / genetics
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Interferon-beta / immunology
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Interferon-beta / metabolism
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Male
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Mice
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Mice, Inbred BALB C
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Mice, Inbred C57BL
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Mice, Knockout
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Myeloid Differentiation Factor 88 / genetics
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Myeloid Differentiation Factor 88 / immunology
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Myeloid Differentiation Factor 88 / metabolism
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Oligonucleotides, Antisense / genetics
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Oligonucleotides, Antisense / immunology*
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Receptors, Interleukin-1 / genetics
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Receptors, Interleukin-1 / immunology
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Receptors, Interleukin-1 / metabolism
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Ribose / genetics
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Ribose / immunology*
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Ribose / metabolism
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Signal Transduction / genetics
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Signal Transduction / immunology
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Toll-Like Receptors / genetics
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Toll-Like Receptors / immunology
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Toll-Like Receptors / metabolism
Substances
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Adaptor Proteins, Signal Transducing
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Adaptor Proteins, Vesicular Transport
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IPS-1 protein, mouse
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Interferon Type I
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Myd88 protein, mouse
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Myeloid Differentiation Factor 88
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Oligonucleotides, Antisense
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Receptors, Interleukin-1
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TICAM-1 protein, mouse
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Toll-Like Receptors
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Ribose
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Interferon-beta
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DNA
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Alanine Transaminase
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Ifih1 protein, mouse
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DEAD-box RNA Helicases
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Interferon-Induced Helicase, IFIH1