Int J Med Sci 2020; 17(12):1840-1853. doi:10.7150/ijms.46339

Research Paper

Comparative Analysis of Global Gene Expression and Complement Components Levels in Umbilical Cord Blood from Preterm and Term Neonates: Implications for Significant Downregulation of Immune Response Pathways related to Prematurity

Dorota Gródecka-Szwajkiewicz1*, Zofia Ulańczyk1*, Edyta Zagrodnik1*, Karolina Łuczkowska1, Dorota Rogińska1, Miłosz P. Kawa1, Iwona Stecewicz1, Krzysztof Safranow2, Przemysław Ustianowski3, Sławomir Szymański4, Bogusław Machaliński1✉

1. Department of General Pathology, Pomeranian Medical University in Szczecin, Szczecin, Poland
2. Department of Biochemistry and Medical Chemistry, Pomeranian Medical University in Szczecin, Szczecin, Poland
3. Department of Perinatology, Obstetrics and Gynecology, Pomeranian Medical University in Szczecin, Szczecin, Poland
4. Department of Obstetrics and Pathology of Pregnancy, Pomeranian Medical University in Szczecin, Szczecin, Poland
*These authors equally contributed to the work.

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Citation:
Gródecka-Szwajkiewicz D, Ulańczyk Z, Zagrodnik E, Łuczkowska K, Rogińska D, Kawa MP, Stecewicz I, Safranow K, Ustianowski P, Szymański S, Machaliński B. Comparative Analysis of Global Gene Expression and Complement Components Levels in Umbilical Cord Blood from Preterm and Term Neonates: Implications for Significant Downregulation of Immune Response Pathways related to Prematurity. Int J Med Sci 2020; 17(12):1840-1853. doi:10.7150/ijms.46339. Available from http://www.medsci.org/v17p1840.htm

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Abstract

Background: Preterm birth is the most frequent cause of neonatal death, but its aetiology remains unclear. It has been suggested that the imbalance of immunological mechanisms responsible for maintaining pregnancy is contributing to preterm birth pathogenesis. We aimed to investigate global gene expression and the levels of several complement system components in umbilical cord blood samples from preterm neonates and compare them to term newborns. We sought to examine how differentially expressed genes could affect various immune-related pathways that are believed to be crucial factors in preterm birth.

Material and methods: We enrolled 27 preterm infants (<37 weeks GA) and 52 term infants (>37 weeks GA), from which umbilical cord blood samples were collected. From these samples, peripheral blood mononuclear cells were isolated and subsequent RNA isolation was performed. We used Affymetrix Human Gene 2.1 ST Array Strip for microarray experiment and DAVID resources for bioinformatics analysis of the obtained data. Concentrations of C2, C3a, C5/C5a, C9, FactorD, Properdin were measured in umbilical cord blood plasma samples using multiplex fluorescent bead-based immunoassays using Luminex technology.

Results: The levels of C3a and C5/5a were significantly elevated in preterm neonates compared to term babies, whereas C9 concentration was evidently increased in term infants. The expression of 250 genes was upregulated at least 2-fold and 3781 genes were downregulated at least 2-fold in preterm neonates in comparison with term infants. Functional annotation analysis revealed that in preterm infants in comparison to term babies there was a significant downregulation of genes encoding several Toll-like receptors, interleukins and genes involved in major signalling pathways (e.g. NF-κB, MAPK, TNF, Notch, JAK) and vital cellular processes (e.g. intracellular signal transduction, protein ubiquitination, protein transport, RNA splicing, DNA-templated transcription).

Conclusions: Preterm birth results in immediate and long-term complications. Our results indicate that infants born prematurely show significant differences in complement components concentration and a downregulation of over 3,000 genes, involved mainly in various immune-related pathways, including innate immune response, phagocytosis and TLR function, when compared to full-term babies. Further studies on larger cohorts are needed to elucidate the role of immunity in prematurity.

Keywords: premature birth, complement system proteins, immunity, gene expression