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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 2

Amniotic fluid reflects tissue- and cell type–specific maturation programs.

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Amniotic fluid reflects tissue- and cell type–specific maturation progra...
(A–C) Predicted relative frequency of Rhesus (A) amniotic fluid, (B) lung, and (C) hippocampus relative to appropriate reference tissue/single-cell collections, using deconvolution. (A) Predicted contribution of specific cell types to amniotic fluid relative to adult human tissue bulk RNA-Seq with GTEx, with only the most frequently detected tissues shown with specific estimates for all rhesus samples by treatment groups. (B) Predicted frequency of rhesus lung cell types, based on human neonatal, child, and adult scRNA-Seq samples. (C) Predicted frequency of Rhesus hippocampus cell-types using human fetal hippocampus cell population scRNA-Seq as a reference. (D) Volcano plot of differentially genes comparing the limma P value and fold change for all genes in term amniotic fluid versus preterm controls. Human term-induced genes from amniotic fluid defined previously are highlighted red (significant) or blue (nonsignificant) (fold ≥ 1.25 and limma P ≤ 0.05 adjusted for treatment year effects). Dashed reference lines mark fold change (vertical) and P value (horizontal) thresholds. (E) UpSet-style plot (marginal intersection counts) indicating overlap of differentially expressed genes in human neonatal versus midgestation hippocampus [Hs], compared with rhesus macaque term versus preterm hippocampus (25) and/or amniotic fluid [Rh]. Genes associated with oligodendrocyte cell identity and myelin sheath formation are highlighted. 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; PTC, preterm control; AT1/2, alveolar type 1/2; OPC, oligodendrocyte precursor cell; InN, inhibitory interneuron; and Non.DG.ExN.2, nondentate gyrus excitatory neuron 2. Data are presented as individual values; boxes represent median, 25th, and 75th percentiles; whiskers extend to ± 1.5 interquartile range. n = 47 animals (lung), 35 animals (amniotic fluid), and 46 animals (hippocampus).

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