scRNA-seq of mouse 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) induced liver injury model

Regulatory T cells (Tregs) were shown to protect from cholestatic liver injury. Here we investigate whether amphiregulin (AREG), a ligand for epithelial growth factor receptor (EGFR) which promotes epithelial repair, mediates innate Treg functions in cholangiopathies. Mice were fed with diet containing 0.1 % of the xenobiotic 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) for 14 days to induce sclerosing cholangitis, followed by a 28-day recovery period (without DDC). Liver mononuclear and epithelial cells were enriched and subjected to single-cell sequencing at six time points: baseline, at days 7 or 14 of DDC treatment, and following a 7-, 14- or 28-day recovery period from 14 days of DDC treatment. Overall design: For our time course, 8-week wild type C57Bl/6 mice were fed with 5058 chow admixed with 0.1% DDC for 14 days and then allowed to recover for an additional 7-28 days (5058 chow without DDC). Liver mononuclear and epithelial cells were enriched and subjected to single-cell sequencing at six time points: baseline, at days 7 or 14 of DDC treatment, and following a 7-, 14- or 28-day recovery period from 14 days of DDC treatment.已有研究证实调节性T细胞（Regulatory T cells, Tregs）可抵御胆汁淤积性肝损伤。本研究旨在探究双调蛋白（amphiregulin, AREG）——一种可促进上皮修复的表皮生长因子受体（epithelial growth factor receptor, EGFR）配体——是否介导了胆管病中的天然Treg功能。我们将小鼠喂食含有0.1%外源性物质3,5-二乙氧羰基-1,4-二氢collidine（DDC）的饲料14天，以诱导硬化性胆管炎，随后设置28天的恢复期（不添加DDC）。分别在6个时间点对富集得到的肝脏单核细胞与上皮细胞进行单细胞测序（single-cell sequencing）：基线期、DDC处理第7天或第14天，以及DDC处理14天后分别恢复7、14或28天的时间点。实验整体设计：本时间序列实验中，8周龄野生型C57Bl/6小鼠被喂食添加0.1% DDC的5058饲料14天，随后更换为不含DDC的5058饲料继续恢复7至28天。对富集得到的肝脏单核细胞与上皮细胞进行单细胞测序，测序时间点共6个：基线期、DDC处理第7天或第14天，以及DDC处理14天后分别恢复7、14或28天的时间点。