Mouse population-guided resequencing reveals that variants in CD44 contribute to acetaminophen-induced liver injury in humans - PubMed (original) (raw)

. 2009 Sep;19(9):1507-15.

doi: 10.1101/gr.090241.108. Epub 2009 May 5.

Paul B Watkins, Stephen Su, Pamela K Ross, David E Harbourt, Ioannis M Stylianou, Gary A Boorman, Mark W Russo, Richard S Sackler, Stephen C Harris, Philip C Smith, Raymond Tennant, Molly Bogue, Kenneth Paigen, Christopher Harris, Tanupriya Contractor, Timothy Wiltshire, Ivan Rusyn, David W Threadgill

Affiliations

Mouse population-guided resequencing reveals that variants in CD44 contribute to acetaminophen-induced liver injury in humans

Alison H Harrill et al. Genome Res. 2009 Sep.

Abstract

Interindividual variability in response to chemicals and drugs is a common regulatory concern. It is assumed that xenobiotic-induced adverse reactions have a strong genetic basis, but many mechanism-based investigations have not been successful in identifying susceptible individuals. While recent advances in pharmacogenetics of adverse drug reactions show promise, the small size of the populations susceptible to important adverse events limits the utility of whole-genome association studies conducted entirely in humans. We present a strategy to identify genetic polymorphisms that may underlie susceptibility to adverse drug reactions. First, in a cohort of healthy adults who received the maximum recommended dose of acetaminophen (4 g/d x 7 d), we confirm that about one third of subjects develop elevations in serum alanine aminotransferase, indicative of liver injury. To identify the genetic basis for this susceptibility, a panel of 36 inbred mouse strains was used to model genetic diversity. Mice were treated with 300 mg/kg or a range of additional acetaminophen doses, and the extent of liver injury was quantified. We then employed whole-genome association analysis and targeted sequencing to determine that polymorphisms in Ly86, Cd44, Cd59a, and Capn8 correlate strongly with liver injury and demonstrated that dose-curves vary with background. Finally, we demonstrated that variation in the orthologous human gene, CD44, is associated with susceptibility to acetaminophen in two independent cohorts. Our results indicate a role for CD44 in modulation of susceptibility to acetaminophen hepatotoxicity. These studies demonstrate that a diverse mouse population can be used to understand and predict adverse toxicity in heterogeneous human populations through guided resequencing.

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Figures

Figure 1.

Figure 1.

Maximum serum ALT fold change measured in human volunteers taking daily oral doses of acetaminophen. The peak ALT fold change over baseline reached over the course of treatment by each subject in the UNC cohort is shown. Subjects were considered responders (N = 34) if peak serum ALT reached greater than 1.5-fold (line) higher than the subject's baseline value.

Figure 2.

Figure 2.

Response to the acute dose of acetaminophen in a panel of mouse strains. (A) Representative photomicrographs (100×) of the hematoxylin and eosin-stained sections of left liver lobe of mice 24 h after dosing with acetaminophen (300 mg/kg). (B) Liver necrosis score (mean ± SE, n = 3–4/strain) in mice treated with acetaminophen (300 mg/kg) for 24 h. (C) Serum ALT levels (mean ± SE) in acetaminophen-treated mice sacrificed 24 h after dosing. (D) Serum ALT levels (mean ± SE) in acetaminophen-treated mice sacrificed 4 h post-dosing. (E) Liver reduced glutathione (ratio between acetaminophen- and vehicle-treated animals in each strain, mean ± SE) 4 h post-dosing. (formula image) Strains with no data. (F) Dose-response to acetaminophen-induced liver injury as measured by ALT release (n = 4/strain, mean ± SE) at 24 h after treatment.

Figure 3.

Figure 3.

Plasma AUC of acetaminophen (mean ± SE) measured across strains for 6 h post-dosing with 50 mg/kg (i.g.) (A) or 300 mg/kg (i.g.) (B) following an overnight fast. Asterisk indicates significant differences between strains by the Tukey post-hoc test.

Figure 4.

Figure 4.

Haplotype association mapping of acetaminophen-induced liver injury in the mouse. Serum ALT at 4 h (A) and 24 h (B) after acetaminophen (300 mg/kg) treatment was used to identify genomic intervals significantly associated with liver injury. Peaks (numbered, see Table 1) indicate a significant log_P_ association score at each 3-SNP marker window. Marker colors indicate chromosome number across the mouse genome.

Figure 5.

Figure 5.

Polymorphisms in CD44 (A–C) and CAPN10 (D–F) associated with susceptibility to acetaminophen-induced liver injury in humans. Data from UNC (A ,D), Purdue Pharma (B ,C) and a combined cohort (C ,F) are shown. Average mean (±SE) serum ALT per genotype is plotted for each matching study day.

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