ABSTRACT

Maternal stress is associated with adverse child health. Breast milk microRNAs encapsulated in extracellular vesicles (EVs) are involved in mother-infant biochemical communication during early-life programming. We leverage the PRogramming of Intergenerational Stress Mechanisms (PRISM) pregnancy cohort to investigate associations between maternal stress and breast milk EV-microRNAs. Lifetime stress and negative life events (NLEs) during pregnancy were assessed using the Life Stressor Checklist-Revised (LSCR) and the Crisis in Family Systems-Revised surveys, respectively. RNA was extracted from breast milk EVs (N = 80; collected 6.1 ± 5.9 weeks postnatally), and microRNAs were profiled using the TaqMan OpenArray Human miRNA panel. Associations between stress scores and detection (yes/no) of 173 microRNAs identified in 20–80% of samples were assessed using logistic regression; associations with expression levels of 205 EV-microRNAs identified in >50% of samples were assessed using linear regression. In adjusted models, detection of 60 and 44 EV-microRNAs was associated with higher LSCR and NLE scores, respectively (p < 0.05). Expression level of 8 and 17 EV-microRNAs was associated with LSCR and NLE scores, respectively, at our a priori criteria of p < 0.05 and |B_regression|>0.2. Enriched KEGG pathways for microRNAs associated with stress scores included fatty acid metabolism and the Hippo signaling pathway. Maternal lifetime stress and NLEs during pregnancy were both associated with detection and expression level of breast milk EV-microRNAs, although associations with microRNA profiles differed between stress measures. Further research is needed to identify biological pathways impacted by associated microRNAs and investigate relationships with child health outcomes.

Abbreviations: EV: extracellular vesicle; PRISM: PRogramming of Intergenerational Stress Mechanisms pregnancy cohort; LSCR: Life Stressor Checklist-Revised survey; NLE: negative life event; CRISYS-R: Crisis in Family Systems-Revised survey; KEGG: Kyoto Encyclopaedia of Genes and Genomes; NYC: New York City; SD: standard deviation; IQR: interquartile range; C_q: relative cycle threshold values; PCA: principal component analysis

摘要

母亲的压力与不利的儿童健康有关。包裹在细胞外囊泡 (EV) 中的母乳 microRNA 在生命早期编程过程中参与母婴生化通讯。我们利用代际压力机制 (PRISM) 妊娠队列规划来研究母体压力与母乳 EV-microRNA 之间的关联。怀孕期间的终生压力和负面生活事件 (NLE) 分别使用修订的生活压力清单 (LSCR) 和修订的家庭系统危机调查进行评估。从母乳 EV（N = 80；产后 6.1 ± 5.9 周收集）中提取 RNA，并使用 TaqMan OpenArray Human miRNA panel 对 microRNA 进行分析。使用逻辑回归评估了在 20-80% 的样本中鉴定出的 173 个 microRNA 的压力评分和检测（是/否）之间的关联；使用线性回归评估了在 >50% 的样本中鉴定出的 205 种 EV-microRNA 表达水平的关联。在调整后的模型中，检测到 60 个和 44 个 EV-microRNA 分别与较高的 LSCR 和 NLE 评分相关（p  < 0.05)。在我们的 先验 标准 p  < 0.05 和 | B_回归|>0.2。与压力评分相关的 microRNA 的丰富 KEGG 通路包括脂肪酸代谢和 Hippo 信号通路。母亲一生的压力和怀孕期间的 NLE 都与母乳 EV-microRNA 的检测和表达水平相关，尽管压力测量与 microRNA 谱的关联不同。需要进一步的研究来确定受相关 microRNA 影响的生物学途径，并调查与儿童健康结果的关系。

缩写： EV：细胞外囊泡；棱镜：代际压力机制妊娠队列的规划；LSCR：生活压力源检查表修订调查；NLE：负面生活事件；CRISYS-R：家庭系统危机修订调查；KEGG：京都基因和基因组百科全书；纽约市：纽约市；SD：标准差；IQR：四分位距；C _q : 相对循环阈值；PCA：主成分分析