Prenatal smoke exposure is a risk factor for impaired lung development in children. Recent studies have indicated that Amphiregulin (AREG), which is a ligand of the epidermal growth factor receptor (EGFR), has a regulatory role in airway epithelial cell differentiation. In this study, we investigated the effect of prenatal smoke exposure on lung epithelial cell differentiation and linked this with AREG-EGFR signaling in 1-day old mouse offspring. Bronchial and alveolar epithelial cell differentiation were assessed by immunohistochemistry. Areg, Egf as well as mRNA expression of specific markers for bronchial and alveolar epithelial cells were assessed by RT-qPCR. Results in neonatal lungs were validated in an AREG-treated 3D mouse lung organoid model. We found that prenatal smoke exposure reduced the number of ciliated cells and the expression of the cilia-related transcription factor Foxj1, whereas it resulted in higher expression of mucus-related transcription factors Spdef and Foxm1 in lung. Moreover, prenatally smoke exposed offspring had higher numbers of alveolar epithelial type II cells (AECII) and lower expression of the AECI-related Pdpn and Gramd2 markers. This was accompanied by higher expression of Areg and lower expression of Egf in prenatal smoke exposed offspring. In bronchial organoids, AREG treatment resulted in fewer ciliated cells and more basal cells when compared non-treated bronchiolar organoids. In alveolar organoids, AREG treatment led to more AECII cells than non-treated AECII cells. Taken together, the observed impaired bronchial and alveolar cell development in prenatally smoke-exposed neonatal offspring may be induced by increased AREG-EGFR signaling.

中文翻译：

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产前烟雾暴露可异常调节小鼠后代的肺上皮细胞分化-AREG诱导的EGFR信号传导的作用。

产前烟雾暴露是儿童肺发育受损的危险因素。最近的研究表明，表皮生长因子受体（EGFR）的配体Amphiregulin（AREG）在气道上皮细胞分化中具有调节作用。在这项研究中，我们调查了产前烟雾暴露对肺上皮细胞分化的影响，并将其与1日龄小鼠后代中的AREG-EGFR信号传导联系起来。通过免疫组织化学评估支气管和肺泡上皮细胞的分化。通过RT-qPCR评估Areg，Egf以及支气管和肺泡上皮细胞特异性标记物的mRNA表达。在AREG处理的3D小鼠肺类器官模型中验证了新生儿肺中的结果。我们发现，产前烟雾暴露减少了纤毛细胞的数量和纤毛相关转录因子Foxj1的表达，而导致粘液相关转录因子Spdef和Foxm1在肺中的表达更高。此外，产前暴露在烟雾中的后代的肺泡上皮II型细胞（AECII）数量更高，而AECI相关的Pdpn和Gramd2标记物的表达则更低。这伴随着产前暴露于烟雾的后代中Areg的高表达和Egf的低表达。与未处理的细支气管类器官相比，在支气管类器官中，AREG处理导致更少的纤毛细胞和更多的基底细胞。在肺泡类器官中，与未处理的AECII细胞相比，AREG处理可导致更多的AECII细胞。在一起