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Cell-Free Fetal DNA Increases Prior to Labor at Term and in a Subset of Preterm Births

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Abstract

Cell-free fetal DNA in the maternal circulation has been associated with the onset of labor at term. Moreover, clinical studies have suggested that cell-free fetal DNA has value to predict pregnancy complications such as spontaneous preterm labor leading to preterm birth. However, a mechanistic link between cell-free fetal DNA and preterm labor and birth has not been established. Herein, using an allogeneic mouse model in which a paternal green fluorescent protein (GFP) can be tracked in the fetuses, we established that cell-free fetal DNA (Egfp) concentrations were higher in late gestation compared to mid-pregnancy and were maintained at increased levels during the onset of labor at term, followed by a rapid decrease after birth. A positive correlation between cell-free fetal DNA concentrations and the number of GFP-positive pups was also observed. The increase in cell-free fetal DNA concentrations prior to labor at term was not linked to a surge in any specific cytokine/chemokine; yet, specific chemokines (i.e., CCL2, CCL7, and CXCL2) increased as gestation progressed and maintained elevated levels in the postpartum period. In addition, cell-free fetal DNA concentrations increased prior to systemic inflammation-induced preterm birth, which was associated with a strong cytokine response in the maternal circulation. However, cell-free fetal DNA concentrations were not increased prior to intra-amniotic inflammation-induced preterm birth, but in this model, a mild inflammatory response was observed in the maternal circulation. Collectively, these findings suggest that an elevation in cell-free fetal DNA concentrations in the maternal circulation precedes the physiological process of labor at term and the pathological process of preterm labor linked with systemic inflammation, but not that associated with intra-amniotic inflammation.

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Acknowledgments

We thank the research assistants from the PRB Perinatal Translational Science Laboratory for their help with carrying out the Luminex assays, and Nicole Ducharme for her help with the DNA extraction and PCR assays. Finally, we thank Derek Miller, MSc, for his critical readings of the manuscript.

Funding

This research was supported, in part, by the Perinatology Research Branch (PRB), Division of Intramural Research, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, US Department of Health and Human Services (NICHD/NIH/DHHS), and, in part, with federal funds from the NICHD/NIH/DHHS under Contract No. HHSN275201300006C. Dr. Roberto Romero has contributed to this work as part of his official duties as an employee of the United States Federal Government. This research was also supported by the Wayne State University Perinatal Initiative in Maternal, Perinatal, and Child Health.

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Authors and Affiliations

  1. Perinatology Research Branch, Division of Obstetrics and Maternal-Fetal Medicine, Division of Intramural Research, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, US Department of Health and Human Services, Bethesda, MD, and Detroit, MI, USA

    Nardhy Gomez-Lopez, Roberto Romero, George Schwenkel, Valeria Garcia-Flores, Bogdan Panaitescu, Aneesha Varrey & Sonia S. Hassan

  2. Department of Obstetrics and Gynecology, Wayne State University School of Medicine, Detroit, MI, USA

    Nardhy Gomez-Lopez, George Schwenkel, Valeria Garcia-Flores, Bogdan Panaitescu, Aneesha Varrey & Sonia S. Hassan

  3. Department of Immunology, Microbiology and Biochemistry, Wayne State University School of Medicine, Detroit, MI, USA

    Nardhy Gomez-Lopez

  4. Department of Obstetrics and Gynecology, University of Michigan, Detroit, MI, USA

    Roberto Romero

  5. Department of Epidemiology and Biostatistics, Michigan State University, East Lansing, MI, USA

    Roberto Romero

  6. Center for Molecular Medicine and Genetics, Wayne State University, Detroit, MI, USA

    Roberto Romero

  7. Detroit Medical Center, Detroit, MI, USA

    Roberto Romero

  8. Department of Obstetrics and Gynecology, Florida International University, Miami, FL, USA

    Roberto Romero

  9. Wayne State University School of Medicine, Detroit, MI, USA

    Fatime Ayoub

  10. Department of Physiology, Wayne State University School of Medicine, Detroit, MI, USA

    Sonia S. Hassan

  11. Vincent Center for Reproductive Biology, Department of Obstetrics and Gynecology, Massachusetts General Hospital, Boston, MA, USA

    Mark Phillippe

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  1. Nardhy Gomez-Lopez
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Electronic supplementary material

Supplementary Figure 1.

Plot showing the maternal plasma concentrations of IL-10 at 12.5 – 19.5 days post coitum (dpc), in labor, and 12 – 36 h postpartum. Each dot represents one maternal plasma sample. Midlines = medians, boxes = interquartile range, whiskers = minimum and maximum range. Red curves indicate trends over time. N = 5 – 11 dams per group. (PNG 62 kb)

High Resolution Image (TIF 910 kb)

Supplementary Figure 2.

Plots showing the negative correlations between plasma concentrations of IL-6 or IL-4 and the number of GFP-positive pups prior to systemic inflammation-induced preterm birth. Spearman correlation coefficients and p-values are displayed. Asterisk (*) indicates p-value obtained using a one-tailed Mann-Whitney U test. N = 8 dams. (PNG 83 kb)

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Supplementary Figure 3.

Plot showing a negative correlation between plasma concentrations of IL-4 and cell-free fetal DNA (Egfp) levels prior to systemic inflammation-induced preterm birth. Spearman correlation coefficients and p-values are displayed. N = 8 dams. (PNG 80 kb)

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Gomez-Lopez, N., Romero, R., Schwenkel, G. et al. Cell-Free Fetal DNA Increases Prior to Labor at Term and in a Subset of Preterm Births. Reprod. Sci. 27, 218–232 (2020). https://doi.org/10.1007/s43032-019-00023-6

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  • DOI: https://doi.org/10.1007/s43032-019-00023-6

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