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 Michael Demarzo, Traci Speed, Zachery R. Reichert, Bernard Kwabi-Addo, Colin J. Henderson, C. Roland Wolf, S Yegnasubramanian, William G Nelson

Research output: Contribution to journalArticle

Abstract

Background: Glutathione S-transferases (GSTs) metabolize drugs and xenobiotics. Yet despite high protein sequence homology, expression of π--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. π--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 π--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, π--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 π--class GSTs may be critical determinants of toxin-induced hepatocyte injury even when not expressed by hepatocytes. Conclusions: By recapitulating human π--class GST expression, hGSTP1+mGstp1/2-/- mice may better model human drug and xenobiotic toxicology.

Original languageEnglish (US)
Article numbere25707
JournalPLoS One
Volume6
Issue number10
DOIs
StatePublished - Oct 11 2011

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acetaminophen
overdose
hepatotoxicity
Acetaminophen
Transferases
Glutathione Transferase
transferases
Liver
Glutathione
Toxicity
glutathione
glutathione transferase
animal models
liver
mice
hepatocytes
Hepatocytes
toxicology
Xenobiotics
xenobiotics

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

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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; Demarzo, Angelo Michael; Speed, Traci; Reichert, Zachery R.; Kwabi-Addo, Bernard; Henderson, Colin J.; Wolf, C. Roland; Yegnasubramanian, S; Nelson, William G.

In: PLoS One, Vol. 6, No. 10, e25707, 11.10.2011.

Research output: Contribution to journalArticle

Vaughn, Matthew P. ; Shinohara, Debika Biswal ; Castagna, Nicole ; Hicks, Jessica L. ; Netto, George ; Demarzo, Angelo Michael ; Speed, Traci ; Reichert, Zachery R. ; Kwabi-Addo, Bernard ; Henderson, Colin J. ; Wolf, C. Roland ; Yegnasubramanian, S ; 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.
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abstract = "Background: Glutathione S-transferases (GSTs) metabolize drugs and xenobiotics. Yet despite high protein sequence homology, expression of π--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. π--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 π--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, π--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 π--class GSTs may be critical determinants of toxin-induced hepatocyte injury even when not expressed by hepatocytes. Conclusions: By recapitulating human π--class GST expression, hGSTP1+mGstp1/2-/- mice may better model human drug and xenobiotic toxicology.",
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AU - Hicks, Jessica L.

AU - Netto, George

AU - Demarzo, Angelo Michael

AU - Speed, Traci

AU - Reichert, Zachery R.

AU - Kwabi-Addo, Bernard

AU - Henderson, Colin J.

AU - Wolf, C. Roland

AU - Yegnasubramanian, S

AU - Nelson, William G

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N2 - Background: Glutathione S-transferases (GSTs) metabolize drugs and xenobiotics. Yet despite high protein sequence homology, expression of π--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. π--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 π--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, π--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 π--class GSTs may be critical determinants of toxin-induced hepatocyte injury even when not expressed by hepatocytes. Conclusions: By recapitulating human π--class GST expression, hGSTP1+mGstp1/2-/- mice may better model human drug and xenobiotic toxicology.

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