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Reduction of liver Fas expression by an antisense oligonucleotide protects mice from fulminant hepatitis

Nature Biotechnology volume 18, pages 862–867 (2000)Cite this article

Abstract

Aberrant apoptosis-mediated cell death is believed to result in a number of different human diseases. For example, excessive apoptosis in the liver can result in fulminant and autoimmune forms of hepatitis. We have explored the possibility that inhibition of Fas expression in mice would reduce the severity of fulminant hepatitis. To do this, we have developed a chemically modified 2′-O-(2-methoxy)ethyl antisense oligonucleotide (ISIS 22023) inhibitor of mouse Fas expression. In tissue culture, this oligonucleotide induced a reduction in Fas mRNA expression that was both concentration- and sequence-specific. In Balb/c mice, dosing with ISIS 22023 reduced Fas mRNA and protein expressions in liver by 90%. The ID50 for this response was 8-10 mg kg−1 daily dosing, and the reduction was highly dependent on oligonucleotide sequence, oligonucleotide concentration in liver, and treatment time. Pretreatment with ISIS 22023 completely protected mice from fulminant hepatitis induced by agonistic Fas antibody, by a mechanism entirely consistent with an oligonucleotide antisense mechanism of action. In addition, oligonucleotide-mediated suppression of Fas expression reduced the severity of acetaminophen-mediated fulminant hepatitis, but was without effect on concanavalin A-mediated hepatitis. Our results demonstrate that 2′-O-(2-methoxy)ethyl containing antisense oligonucleotides targeting Fas can exert in vivo pharmacological activity in liver, and suggest that oligonucleotide inhibitors of Fas may be useful in the treatment of human liver disease.

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Figure 1: Effect of antisense oligonucleotides on Fas mRNA expression in tissue culture and mouse liver.
Figure 2: Effect of antisense oligonucleotides on Fas protein expression in mouse liver.
Figure 3: Effects of antisense oligonucleotide-mediated Fas inhibition in liver on the progression of agonistic Fas antibody-induced fulminant hepatitis.
Figure 4: Effect of antisense oligonucleotides on AAP-induced Fas expression in liver and circulating ALT activity.
Figure 5: Effect of antisense oligonucleotide-mediated Fas inhibition in mouse liver on AAP-induced hepatocyte apoptosis.

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Acknowledgements

We thank Boyd Conklin and Henri Sasmor for the synthesis of oligonucleotides.

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Authors and Affiliations

  1. Department of Pharmacology, Isis Pharmaceuticals, Inc., Carlsbad, 92008, CA

    Hong Zhang, Jesse Cook, Jeffrey Nickel, Rosie Yu, Kimberly Stecker, Kathleen Myers & Nicholas M. Dean

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  1. Hong Zhang
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Correspondence to Hong Zhang.

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Zhang, H., Cook, J., Nickel, J. et al. Reduction of liver Fas expression by an antisense oligonucleotide protects mice from fulminant hepatitis. Nat Biotechnol 18, 862–867 (2000). https://doi.org/10.1038/78475

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