Center for Advanced Studies and Technology (CAST), "G. d'Annunzio" University of Chieti-Pescara, Chieti 66100, Italy.
Department of Psychological, Health and Territorial Sciences, "G. d'Annunzio" University of Chieti-Pescara, 66100 Chieti, Italy.
Curr Pharm Des. 2024;30(25):1995-2006. doi: 10.2174/0113816128305232240607084420.
Smoking during pregnancy has been linked to adverse health outcomes in offspring, but the underlying mechanisms are not fully understood. To date, the effect of maternal smoking has been tested in primary tissues and animal models, but the scarcity of human tissues limits experimental studies. Evidence regarding smoking-related molecular alteration and gene expression profiles in stem cells is still lacking.
We developed a cell culture model of human amniotic fluid stem cells (hAFSCs) of nicotine (NIC) exposure to examine the impact of maternal smoking on epigenetic alterations of the fetus.
NIC 0.1 μM (equivalent to "light" smoking, i.e., 5 cigarettes/day) did not significantly affect cell viability; however, significant alterations in DNA methylation and N6-methyladenosine (m6A) RNA methylation in hAFSCs occurred. These epigenetic changes may influence the gene expression and function of hAFSCs. Furthermore, NIC exposure caused time-dependent alterations of the expression of pluripotency genes and cell surface markers, suggesting enhanced cell stemness and impaired differentiation potential. Furthermore, NICtreated cells showed reduced mRNA levels of key adipogenic markers and hypomethylation of the promoter region of the imprinted gene during adipogenic differentiation, potentially suppressing adipo/lipogenesis. Differential expression of 16 miRNAs, with predicted target genes involved in various metabolic pathways and linked to pathological conditions, including cognitive delay and fetal growth retardation, has been detected.
Our findings highlight multi-level effects of NIC on hAFSCs, including epigenetic modifications, altered gene expression, and impaired cellular differentiation, which may contribute to long-term consequences of smoking in pregnancy and its potential impact on offspring health and development.
孕期吸烟与后代不良健康结局有关，但其中的潜在机制尚不完全清楚。迄今为止，人们已经在原代组织和动物模型中检验了母亲吸烟的影响，但人类组织的稀缺性限制了实验研究。关于吸烟相关的分子改变和干细胞中的基因表达谱的证据仍然缺乏。
我们开发了一种人羊水干细胞（hAFSCs）的尼古丁（NIC）暴露细胞培养模型，以研究母亲吸烟对胎儿表观遗传改变的影响。
NIC 0.1 μM（相当于“轻度”吸烟，即每天 5 支香烟）对细胞活力没有显著影响；然而，hAFSCs 中的 DNA 甲基化和 N6-甲基腺苷（m6A）RNA 甲基化发生了显著改变。这些表观遗传变化可能影响 hAFSCs 的基因表达和功能。此外，NIC 暴露导致多能性基因和细胞表面标志物的表达出现时间依赖性改变，提示增强了细胞干性和受损的分化潜能。此外，NIC 处理的细胞在成脂分化过程中表现出关键成脂标志物的 mRNA 水平降低和印迹基因启动子区域的低甲基化，可能抑制脂肪形成/脂肪生成。检测到 16 个 miRNA 的差异表达，其预测的靶基因参与各种代谢途径，并与认知延迟和胎儿生长迟缓等病理状况有关。
我们的研究结果强调了 NIC 对 hAFSCs 的多水平影响，包括表观遗传修饰、基因表达改变和细胞分化受损，这可能导致妊娠期间吸烟的长期后果及其对后代健康和发育的潜在影响。
