Humanizing π-class glutathione S-transferase regulation in a mouse model alters liver toxicity in response to acetaminophen overdose

Matthew P. Vaughn, Debika Biswal Shinohara, Nicole Castagna, Jessica L. Hicks, George Netto, Angelo M. De Marzo, Traci J. Speed, Zachery R. Reichert, Bernard Kwabi-Addo, Colin J. Henderson, C. Roland Wolf, Srinivasan Yegnasubramanian, William G. Nelson

    Research output: Contribution to journalArticle

    10 Citations (Scopus)

    Abstract

    Background: Glutathione S-transferases (GSTs) metabolize drugs and xenobiotics. Yet despite high protein sequence homology, expression of p-class GSTs, the most abundant of the enzymes, varies significantly between species. In mouse liver, hepatocytes exhibit high mGstp expression, while in human liver, hepatocytes contain little or no hGSTP1 mRNA or hGSTP1 protein. pi-class GSTs are known to be critical determinants of liver responses to drugs and toxins: when treated with high doses of acetaminophen, mGstp1/2+/+ mice suffer marked liver damage, while mGstp1/2-/- mice escape liver injury.

    Methodology/Principal Findings: To more faithfully model the contribution of p-class GSTs to human liver toxicology, we introduced hGSTP1, with its exons, introns, and flanking sequences, into the germline of mice carrying disrupted mGstp genes. In the resultant hGSTP1+ mGstp1/2-/- strain, pi-class GSTs were regulated differently than in wild-type mice. In the liver, enzyme expression was restricted to bile duct cells, Kupffer cells, macrophages, and endothelial cells, reminiscent of human liver, while in the prostate, enzyme production was limited to basal epithelial cells, reminiscent of human prostate. The human patterns of hGSTP1 transgene regulation were accompanied by human patterns of DNA methylation, with bisulfite genomic sequencing revealing establishment of an unmethylated CpG island sequence encompassing the gene promoter. Unlike wild-type or mGstp1/2-/- mice, when hGSTP1+ mGstp1/2-/- mice were overdosed with acetaminophen, liver tissues showed limited centrilobular necrosis, suggesting that pi-class GSTs may be critical determinants of toxin-induced hepatocyte injury even when not expressed by hepatocytes.

    Conclusions: By recapitulating human p-class GST expression, hGSTP1+ mGstp1/2-/- mice may better model human drug and xenobiotic toxicology.

    Original languageEnglish
    Article numbere25707
    Pages (from-to)-
    Number of pages10
    JournalPLoS ONE
    Volume6
    Issue number10
    DOIs
    Publication statusPublished - 2011

    Keywords

    • RAT CHEMICAL HEPATOCARCINOGENESIS
    • ISLAND DNA HYPERMETHYLATION
    • GSTP1 CPG ISLAND
    • HEPATOCELLULAR-CARCINOMA
    • TERMINAL KINASE
    • TRANSCRIPTIONAL ACTIVATION
    • PROMOTER HYPERMETHYLATION
    • GENE
    • MICE
    • JUN

    Cite this

    Vaughn, M. P., Shinohara, D. B., Castagna, N., Hicks, J. L., Netto, G., De Marzo, A. M., ... Nelson, W. G. (2011). Humanizing π-class glutathione S-transferase regulation in a mouse model alters liver toxicity in response to acetaminophen overdose. PLoS ONE, 6(10), -. [e25707]. https://doi.org/10.1371/journal.pone.0025707
    Vaughn, Matthew P. ; Shinohara, Debika Biswal ; Castagna, Nicole ; Hicks, Jessica L. ; Netto, George ; De Marzo, Angelo M. ; Speed, Traci J. ; Reichert, Zachery R. ; Kwabi-Addo, Bernard ; Henderson, Colin J. ; Wolf, C. Roland ; Yegnasubramanian, Srinivasan ; Nelson, William G. / Humanizing π-class glutathione S-transferase regulation in a mouse model alters liver toxicity in response to acetaminophen overdose. In: PLoS ONE. 2011 ; Vol. 6, No. 10. pp. -.
    @article{b6d7aca4090941c7a2b1e08c72bd94db,
    title = "Humanizing π-class glutathione S-transferase regulation in a mouse model alters liver toxicity in response to acetaminophen overdose",
    abstract = "Background: Glutathione S-transferases (GSTs) metabolize drugs and xenobiotics. Yet despite high protein sequence homology, expression of p-class GSTs, the most abundant of the enzymes, varies significantly between species. In mouse liver, hepatocytes exhibit high mGstp expression, while in human liver, hepatocytes contain little or no hGSTP1 mRNA or hGSTP1 protein. pi-class GSTs are known to be critical determinants of liver responses to drugs and toxins: when treated with high doses of acetaminophen, mGstp1/2+/+ mice suffer marked liver damage, while mGstp1/2-/- mice escape liver injury.Methodology/Principal Findings: To more faithfully model the contribution of p-class GSTs to human liver toxicology, we introduced hGSTP1, with its exons, introns, and flanking sequences, into the germline of mice carrying disrupted mGstp genes. In the resultant hGSTP1+ mGstp1/2-/- strain, pi-class GSTs were regulated differently than in wild-type mice. In the liver, enzyme expression was restricted to bile duct cells, Kupffer cells, macrophages, and endothelial cells, reminiscent of human liver, while in the prostate, enzyme production was limited to basal epithelial cells, reminiscent of human prostate. The human patterns of hGSTP1 transgene regulation were accompanied by human patterns of DNA methylation, with bisulfite genomic sequencing revealing establishment of an unmethylated CpG island sequence encompassing the gene promoter. Unlike wild-type or mGstp1/2-/- mice, when hGSTP1+ mGstp1/2-/- mice were overdosed with acetaminophen, liver tissues showed limited centrilobular necrosis, suggesting that pi-class GSTs may be critical determinants of toxin-induced hepatocyte injury even when not expressed by hepatocytes.Conclusions: By recapitulating human p-class GST expression, hGSTP1+ mGstp1/2-/- mice may better model human drug and xenobiotic toxicology.",
    keywords = "RAT CHEMICAL HEPATOCARCINOGENESIS, ISLAND DNA HYPERMETHYLATION, GSTP1 CPG ISLAND, HEPATOCELLULAR-CARCINOMA, TERMINAL KINASE, TRANSCRIPTIONAL ACTIVATION, PROMOTER HYPERMETHYLATION, GENE, MICE, JUN",
    author = "Vaughn, {Matthew P.} and Shinohara, {Debika Biswal} and Nicole Castagna and Hicks, {Jessica L.} and George Netto and {De Marzo}, {Angelo M.} and Speed, {Traci J.} and Reichert, {Zachery R.} and Bernard Kwabi-Addo and Henderson, {Colin J.} and Wolf, {C. Roland} and Srinivasan Yegnasubramanian and Nelson, {William G.}",
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    Vaughn, MP, Shinohara, DB, Castagna, N, Hicks, JL, Netto, G, De Marzo, AM, Speed, TJ, Reichert, ZR, Kwabi-Addo, B, Henderson, CJ, Wolf, CR, Yegnasubramanian, S & Nelson, WG 2011, 'Humanizing π-class glutathione S-transferase regulation in a mouse model alters liver toxicity in response to acetaminophen overdose', PLoS ONE, vol. 6, no. 10, e25707, pp. -. https://doi.org/10.1371/journal.pone.0025707

    Humanizing π-class glutathione S-transferase regulation in a mouse model alters liver toxicity in response to acetaminophen overdose. / Vaughn, Matthew P.; Shinohara, Debika Biswal; Castagna, Nicole; Hicks, Jessica L.; Netto, George; De Marzo, Angelo M.; Speed, Traci J.; Reichert, Zachery R.; Kwabi-Addo, Bernard; Henderson, Colin J.; Wolf, C. Roland; Yegnasubramanian, Srinivasan; Nelson, William G.

    In: PLoS ONE, Vol. 6, No. 10, e25707, 2011, p. -.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Humanizing π-class glutathione S-transferase regulation in a mouse model alters liver toxicity in response to acetaminophen overdose

    AU - Vaughn, Matthew P.

    AU - Shinohara, Debika Biswal

    AU - Castagna, Nicole

    AU - Hicks, Jessica L.

    AU - Netto, George

    AU - De Marzo, Angelo M.

    AU - Speed, Traci J.

    AU - Reichert, Zachery R.

    AU - Kwabi-Addo, Bernard

    AU - Henderson, Colin J.

    AU - Wolf, C. Roland

    AU - Yegnasubramanian, Srinivasan

    AU - Nelson, William G.

    PY - 2011

    Y1 - 2011

    N2 - Background: Glutathione S-transferases (GSTs) metabolize drugs and xenobiotics. Yet despite high protein sequence homology, expression of p-class GSTs, the most abundant of the enzymes, varies significantly between species. In mouse liver, hepatocytes exhibit high mGstp expression, while in human liver, hepatocytes contain little or no hGSTP1 mRNA or hGSTP1 protein. pi-class GSTs are known to be critical determinants of liver responses to drugs and toxins: when treated with high doses of acetaminophen, mGstp1/2+/+ mice suffer marked liver damage, while mGstp1/2-/- mice escape liver injury.Methodology/Principal Findings: To more faithfully model the contribution of p-class GSTs to human liver toxicology, we introduced hGSTP1, with its exons, introns, and flanking sequences, into the germline of mice carrying disrupted mGstp genes. In the resultant hGSTP1+ mGstp1/2-/- strain, pi-class GSTs were regulated differently than in wild-type mice. In the liver, enzyme expression was restricted to bile duct cells, Kupffer cells, macrophages, and endothelial cells, reminiscent of human liver, while in the prostate, enzyme production was limited to basal epithelial cells, reminiscent of human prostate. The human patterns of hGSTP1 transgene regulation were accompanied by human patterns of DNA methylation, with bisulfite genomic sequencing revealing establishment of an unmethylated CpG island sequence encompassing the gene promoter. Unlike wild-type or mGstp1/2-/- mice, when hGSTP1+ mGstp1/2-/- mice were overdosed with acetaminophen, liver tissues showed limited centrilobular necrosis, suggesting that pi-class GSTs may be critical determinants of toxin-induced hepatocyte injury even when not expressed by hepatocytes.Conclusions: By recapitulating human p-class GST expression, hGSTP1+ mGstp1/2-/- mice may better model human drug and xenobiotic toxicology.

    AB - Background: Glutathione S-transferases (GSTs) metabolize drugs and xenobiotics. Yet despite high protein sequence homology, expression of p-class GSTs, the most abundant of the enzymes, varies significantly between species. In mouse liver, hepatocytes exhibit high mGstp expression, while in human liver, hepatocytes contain little or no hGSTP1 mRNA or hGSTP1 protein. pi-class GSTs are known to be critical determinants of liver responses to drugs and toxins: when treated with high doses of acetaminophen, mGstp1/2+/+ mice suffer marked liver damage, while mGstp1/2-/- mice escape liver injury.Methodology/Principal Findings: To more faithfully model the contribution of p-class GSTs to human liver toxicology, we introduced hGSTP1, with its exons, introns, and flanking sequences, into the germline of mice carrying disrupted mGstp genes. In the resultant hGSTP1+ mGstp1/2-/- strain, pi-class GSTs were regulated differently than in wild-type mice. In the liver, enzyme expression was restricted to bile duct cells, Kupffer cells, macrophages, and endothelial cells, reminiscent of human liver, while in the prostate, enzyme production was limited to basal epithelial cells, reminiscent of human prostate. The human patterns of hGSTP1 transgene regulation were accompanied by human patterns of DNA methylation, with bisulfite genomic sequencing revealing establishment of an unmethylated CpG island sequence encompassing the gene promoter. Unlike wild-type or mGstp1/2-/- mice, when hGSTP1+ mGstp1/2-/- mice were overdosed with acetaminophen, liver tissues showed limited centrilobular necrosis, suggesting that pi-class GSTs may be critical determinants of toxin-induced hepatocyte injury even when not expressed by hepatocytes.Conclusions: By recapitulating human p-class GST expression, hGSTP1+ mGstp1/2-/- mice may better model human drug and xenobiotic toxicology.

    KW - RAT CHEMICAL HEPATOCARCINOGENESIS

    KW - ISLAND DNA HYPERMETHYLATION

    KW - GSTP1 CPG ISLAND

    KW - HEPATOCELLULAR-CARCINOMA

    KW - TERMINAL KINASE

    KW - TRANSCRIPTIONAL ACTIVATION

    KW - PROMOTER HYPERMETHYLATION

    KW - GENE

    KW - MICE

    KW - JUN

    U2 - 10.1371/journal.pone.0025707

    DO - 10.1371/journal.pone.0025707

    M3 - Article

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    VL - 6

    SP - -

    JO - PLoS ONE

    JF - PLoS ONE

    SN - 1932-6203

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    M1 - e25707

    ER -