Identifying TCDD-resistance genes via murine and rat comparative genomics and transcriptomics
Authors: Prokopec Stephenie D. , Lu Aileen , Lee Sandy Che-Eun S. , Yao Cindy Q. , Sun Ren X. , Watson John D. , de Borja Richard , Wong Ada , Sam Michelle , Zuzarte Philip , McPherson John D. , Okey Allan B. , Pohjanvirta Raimo , Boutros Paul C. 
Published: April 2019

The aryl hydrocarbon receptor (AHR) mediates many of the toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). However, the AHR alone is insufficient to explain the widely different outcomes among organisms. Attempts to identify unknown factor(s) have been confounded by genetic variability of model organisms. Here, we evaluated three transgenic mouse lines, each expressing a different rat AHR isoform (rWT, DEL, and INS), as well as C57BL/6 and DBA/2 mice. We supplement these with whole-genome sequencing and transcriptomic analyses of the corresponding rat models: Long-Evans (L-E) and Han/Wistar (H/W) rats. These integrated multi-species genomic and transcriptomic data were used to identify genes associated with TCDD-response phenotypes.

We identified several genes that show consistent transcriptional changes in both transgenic mice and rats. HepaticPxdc1was significantly repressed by TCDD in C57BL/6, rWT mice, and in L-E rat. Three genes demonstrated different AHRE-1 (full) motif occurrences within their promoter regions:Cxxc5had fewer occurrences in H/W, as compared with L-E;Sugp1andHgfac(in either L-E or H/W respectively). These genes also showed different patterns of mRNA abundance across strains.

The AHR isoform explains much of the transcriptional variability: up to 50% of genes with altered mRNA abundance following TCDD exposure are associated with a single AHR isoform (30% and 10% unique to DEL and rWT respectively following 500 μg/kg TCDD). Genomic and transcriptomic evidence allowed identification of genes potentially involved in phenotypic outcomes:Pxdc1had differential mRNA abundance by phenotype;Cxxc5had altered AHR binding sites and differential mRNA abundance.

Author Summary

Environmental contaminants such as dioxins cause many toxic responses, anything from chloracne (common in humans) to death. These toxic responses are mostly regulated by theAhr, a ligand-activated transcription factor with roles in drug metabolism and immune responses, however other contributing factors remain unclear. Studies are complicated by the underlying genetic heterogeneity of model organisms. Our team evaluated a number of mouse and rat models, including two strains of mouse, two strains of rat and three transgenic mouse lines which differ only at theAhrlocus, that present widely different sensitivities to the most potent dioxin: 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD). We identified a number of changes to gene expression that were associated with different toxic responses. We then contrasted these findings with results from whole-genome sequencing of the H/W and L-E rats and found some key genes, such asCxxc5andMafb, which might contribute to TCDD toxicity. These transcriptomic and genomic datasets will provide a valuable resource for future studies into the mechanisms of dioxin toxicities.