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RNA interference targeting Fas protects mice from fulminant hepatitis

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Abstract

RNA interference (RNAi) is a powerful tool to silence gene expression post-transcriptionally1. However, its potential to treat or prevent disease remains unproven. Fas-mediated apoptosis is implicated in a broad spectrum of liver diseases, where inhibiting hepatocyte death is life-saving2. We investigated the in vivo silencing effect of small interfering RNA (siRNA) duplexes targeting the gene Fas (also known as Tnfrsf6), encoding the Fas receptor, to protect mice from liver failure and fibrosis in two models of autoimmune hepatitis. Intravenous injection of Fas siRNA specifically reduced Fas mRNA levels and expression of Fas protein in mouse hepatocytes, and the effects persisted without diminution for 10 days. Hepatocytes isolated from mice treated with Fas siRNA were resistant to apoptosis when exposed to Fas-specific antibody or co-cultured with concanavalin A (ConA)-stimulated hepatic mononuclear cells. Treatment with Fas siRNA 2 days before ConA challenge abrogated hepatocyte necrosis and inflammatory infiltration and markedly reduced serum concentrations of transaminases. Administering Fas siRNA beginning one week after initiating weekly ConA injections protected mice from liver fibrosis. In a more fulminant hepatitis induced by injecting agonistic Fas-specific antibody, 82% of mice treated with siRNA that effectively silenced Fas survived for 10 days of observation, whereas all control mice died within 3 days. Silencing Fas expression with RNAi holds therapeutic promise to prevent liver injury by protecting hepatocytes from cytotoxicity.

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Figure 1: Injection of siRNA duplex efficiently silences Fas gene expression in mouse hepatocytes.
Figure 2: In vivo treatment with Fas siRNA protects mouse hepatocytes from Fas-mediated apoptosis and cytotoxic lysis by ConA-activated hepatic mononuclear cells.
Figure 3: Fas gene silencing protects mice from fulminant hepatitis and hepatic fibrosis.

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  • 14 February 2003

    This was incorrect in AOP version but corrected in print. Changed one word in the legend, as per the note.

Notes

  1. NOTE: In the version of this article initially published online, the Fig. 1 legend contained an error. The third sentence of (b) should read, "...housekeeping genes (L32 and Gapd) were unaffected..." This mistake has been corrected for the HTML and print versions of the article.

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Acknowledgements

This work was supported by US National Institutes of Health grants AI42510 and AI45406 (J.L.), AI49792 and AI45306 (P.S.), American Foundation for AIDS Research (AmfAR) grants 70589-32-RF (S.L.) and 70540-30-RF (N.I.), Guangdong Provincial Natural Science Foundation grants 980098 (E.S.) and 001359 (J.M.), and National Natural Scientific Foundation of China grants 39670705 (J.C.), 39700135 and 39970718 (J.M.).

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Correspondence to Judy Lieberman.

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Song, E., Lee, SK., Wang, J. et al. RNA interference targeting Fas protects mice from fulminant hepatitis. Nat Med 9, 347–351 (2003). https://doi.org/10.1038/nm828

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