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Fetal maturation revealed by amniotic fluid cell-free transcriptome in rhesus macaques
Augusto F. Schmidt, Daniel J. Schnell, Kenneth P. Eaton, Kashish Chetal, Paranthaman S. Kannan, Lisa A. Miller, Claire A. Chougnet, Daniel T. Swarr, Alan H. Jobe, Nathan Salomonis, Beena D. Kamath-Rayne
Augusto F. Schmidt, Daniel J. Schnell, Kenneth P. Eaton, Kashish Chetal, Paranthaman S. Kannan, Lisa A. Miller, Claire A. Chougnet, Daniel T. Swarr, Alan H. Jobe, Nathan Salomonis, Beena D. Kamath-Rayne
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Research Article Reproductive biology

Fetal maturation revealed by amniotic fluid cell-free transcriptome in rhesus macaques

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Abstract

Accurate estimate of fetal maturity could provide individualized guidance for delivery of complicated pregnancies. However, current methods are invasive, have low accuracy, and are limited to fetal lung maturation. To identify diagnostic gestational biomarkers, we performed transcriptomic profiling of lung and brain, as well as cell-free RNA from amniotic fluid of preterm and term rhesus macaque fetuses. These data identify potentially new and prior-associated gestational age differences in distinct lung and neuronal cell populations when compared with existing single-cell and bulk RNA-Seq data. Comparative analyses found hundreds of genes coincidently induced in lung and amniotic fluid, along with dozens in brain and amniotic fluid. These data enable creation of computational models that accurately predict lung compliance from amniotic fluid and lung transcriptome of preterm fetuses treated with antenatal corticosteroids. Importantly, antenatal steroids induced off-target gene expression changes in the brain, impinging upon synaptic transmission and neuronal and glial maturation, as this could have long-term consequences on brain development. Cell-free RNA in amniotic fluid may provide a substrate of global fetal maturation markers for personalized management of at-risk pregnancies.

Authors

Augusto F. Schmidt, Daniel J. Schnell, Kenneth P. Eaton, Kashish Chetal, Paranthaman S. Kannan, Lisa A. Miller, Claire A. Chougnet, Daniel T. Swarr, Alan H. Jobe, Nathan Salomonis, Beena D. Kamath-Rayne

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Figure 1

Evaluation of antenatal corticosteroids across tissues and amniotic fluid during rhesus gestation.

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Evaluation of antenatal corticosteroids across tissues and amniotic flui...
(A) Study design for treatment and C-section of pregnant rhesus macaque females using indicated antenatal corticosteroid dosing. Amniotic fluid and brain (hippocampus) and lung tissue were collected from each fetus from either preterm (127–136 days) or term (156 and 157 days) gestation and analyzed by bulk RNA-Seq analysis. (B) PCA of all genes, following removal of strongly sex-associated genes and NOISeq batch-effect correction, for amniotic fluid, brain, and lungs (mean TPM ≥ 1). (C) Heatmap of the top most-specific marker genes for lung RNA-Seq for each treatment. Expression values were calculated as log2 fold changes relative to the mean of each row. The top GO-Elite Gene Ontology enrichment results are denoted to the left of each cluster, along with corresponding Fischer’s exact test enrichment P values. B060, i.m. betamethasone-acetate 0.06 mg/kg × 1 dose; B125, i.m. betamethasone-acetate 0.125 mg/kg × 1 dose; C1x, i.m. Celestone (betamethasone-acetate + betamethasone-phosphate) 0.25 mg/kg × 1 dose; C2x, i.m. Celestone 0.25 mg/kg × 2 doses; POB, oral betamethasone-phosphate 0.15 mg/kg × 3 doses; POD, oral dexamethasone-phosphate 0.15 mg/kg × 3 doses; and PTC, preterm control. n = 47 animals for lung; 35 animals for amniotic fluid and 46 animals for hippocampus.

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