Patterns of Maternal Neutrophil Gene Expression at 30 Weeks of Gestation, but Not DNA Methylation, Distinguish Mild from Severe Preeclampsia.
Patterns of Maternal Neutrophil Gene Expression at 30 Weeks of Gestation, but Not DNA Methylation, Distinguish Mild from Severe Preeclampsia.
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DOI：
10.3390/ijms222312876
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发表时间：
2021-11-28
影响因子：
5.6
通讯作者：
Strauss JF 3rd
中科院分区：
文献类型：
--
作者：
Walsh SW;Al Dulaimi M;Archer KJ;Strauss JF 3rd
在先兆子痫妇女中，中性粒细胞被激活并广泛浸润血管。为了鉴定有助于中性粒细胞活化和浸润的基因，我们分析了正常孕妇和先兆子痫妇女循环中性粒细胞的转录组。在妊娠30周时收集中性粒细胞，分离RNA和DNA用于RNA测序和5-羟甲基胞嘧啶（5-hmC）测序，作为中性粒细胞DNA甲基化动态变化的指标。正常妊娠的妇女在足月时继续发展为轻度先兆子痫，具有最独特的表达基因（697），其中325个基因本体通路上调，许多与中性粒细胞活化和功能相关。患有严重先兆子痫的早产妇女几乎没有上调或下调的途径。聚类分析显示，重度子痫前期妇女的基因表达与正常妊娠妇女的基因表达呈反镜像，而轻度子痫前期妇女的基因表达与两者有显著差异。DNA甲基化标记是基因表达的关键调节因子，通过10 - 11易位（泰特）酶的作用被去除，所述酶氧化5-甲基胞嘧啶（5 mCs），导致DNA甲基化的基因座特异性逆转。5-hmC的DNA测序显示三组之间没有差异。全基因组DNA甲基化显示循环中性粒细胞中的水平极低，表明它们是去甲基化的。总的来说，这些数据表明，中性粒细胞基因表达谱可以区分不同的先兆子痫表型，在轻度先兆子痫的情况下，基因表达的改变发生在临床症状出现之前。这些发现为进一步评价中性粒细胞转录组作为先兆子痫表型的生物标志物奠定了基础。循环中性粒细胞DNA甲基化的变化似乎并不介导轻度或重度先兆子痫基因表达的差异模式。
Neutrophils are activated and extensively infiltrate blood vessels in preeclamptic women. To identify genes that contribute to neutrophil activation and infiltration, we analyzed the transcriptomes of circulating neutrophils from normal pregnant and preeclamptic women. Neutrophils were collected at 30 weeks’ gestation and RNA and DNA were isolated for RNA sequencing and 5-hydroxy-methylcytosine (5-hmC) sequencing as an index of dynamic changes in neutrophil DNA methylation. Women with normal pregnancy who went on to develop mild preeclampsia at term had the most uniquely expressed genes (697) with 325 gene ontology pathways upregulated, many related to neutrophil activation and function. Women with severe preeclampsia who delivered prematurely had few pathways up- or downregulated. Cluster analysis revealed that gene expression in women with severe preeclampsia was an inverse mirror image of gene expression in normal pregnancy, while gene expression in women who developed mild preeclampsia was remarkably different from both. DNA methylation marks, key regulators of gene expression, are removed by the action of ten-eleven translocation (TET) enzymes, which oxidize 5-methylcytosines (5mCs), resulting in locus-specific reversal of DNA methylation. DNA sequencing for 5-hmC revealed no differences among the three groups. Genome-wide DNA methylation revealed extremely low levels in circulating neutrophils suggesting they are de-methylated. Collectively, these data demonstrate that neutrophil gene expression profiles can distinguish different preeclampsia phenotypes, and in the case of mild preeclampsia, alterations in gene expression occur well before clinical symptoms emerge. These findings serve as a foundation for further evaluation of neutrophil transcriptomes as biomarkers of preeclampsia phenotypes. Changes in DNA methylation in circulating neutrophils do not appear to mediate differential patterns of gene expression in either mild or severe preeclampsia.