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. 2022 Apr 28;23(9):4924.
doi: 10.3390/ijms23094924.

Gene Expression of Pregnancy Neutrophils Differs for Protease versus Lipopolysaccharide Stimulation

Scott W Walsh  1 , Marwah Al Dulaimi  1 , Jerome F Strauss 3rd  1
Affiliations

Gene Expression of Pregnancy Neutrophils Differs for Protease versus Lipopolysaccharide Stimulation

Scott W Walsh et al. Int J Mol Sci. .

Abstract

Neutrophils, which extensively infiltrate maternal systemic blood vessels in preeclampsia, express protease-activated receptor 1 (PAR-1) but only during pregnancy. Neutrophils are generally considered to be non-specific in their response, but the pregnancy-specific expression of PAR-1 could result in a gene expression profile unique to pregnancy, which could help explain why the maternal inflammatory response in preeclampsia is systemic rather than localized. We sought to determine if gene expression of pregnancy neutrophils would differ if stimulated by a protease versus bacterial lipopolysaccharide (LPS). We isolated neutrophils from normal pregnant women at 30 weeks' gestation and cultured them with elastase or LPS. We used elastase because it is a protease elevated in women with preeclampsia, and it activates pregnancy neutrophils via PAR-1. RNA was isolated from the neutrophils for sequencing of the transcriptomes. We discovered many differences in the gene expression profiles. For example, exposure to elastase resulted in three times more uniquely expressed genes than LPS, and the number of significantly differentially upregulated and downregulated genes was greater for elastase. Analysis of canonical pathways revealed similarities for innate immunity but also differences. LPS treatment enriched more pathways, but elastase activated more genes in each pathway. Elastase treatment enriched the MAPK signaling pathway, whereas LPS did not. This is significant because MAPK is a key mediator of transcriptional responses. These findings indicate that protease stimulation of pregnancy neutrophils results in a different profile than stimulation with LPS, which may help explain why the sterile inflammatory response of preeclampsia is systemic and unique to pregnancy.

Keywords: elastase; gene expression; lipopolysaccharide; neutrophils; preeclampsia; pregnancy; protease-activated receptor 1.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Co-expression Venn diagram. Pregnancy neutrophils were obtained at approximately 30 weeks’ gestation and treated with elastase or lipopolysaccharide (LPS). Elastase treatment resulted in 3 times more uniquely expressed genes than LPS. A total of 299 were uniquely expressed with elastase as compared to 96 for LPS.
Figure 2
Figure 2
Cluster analysis heatmap of differentially expressed genes with high (red) expression levels and genes with low (green) expression levels. Cluster analysis of high and low expressed genes revealed significant differences. Elastase and LPS were clearly different from control, but they were also clearly different from each other.
Figure 3
Figure 3
Number of significantly differentially regulated genes (DEG) either upregulated or downregulated by treatments. Elastase upregulated approximately 30% more genes than LPS and downregulated approximately 75% more genes than LPS. Overall, elastase differently regulated more genes than LPS.
Figure 4
Figure 4
Volcano plot for significantly differentially regulated genes comparing elastase with LPS. Elastase upregulated 66 more genes than LPS and downregulated 221 more genes than LPS.
See this image and copyright information in PMC

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