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| Status |
Public on Feb 20, 2021 |
| Title |
Endometrium on-a-chip reveals insulin- and glucose-induced alterations in the transcriptome |
| Organism |
Bos taurus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
The molecular interactions between the maternal environment and the developing embryo that are key for early pregnancy success are known to be influenced by factors such as maternal metabolic status. Our understanding of the mechanism(s) through which these individual nutritional stressors alter endometrial function and the in utero environment for early pregnancy success is, however, limited. Here we report, for the first time, the use of an endometrium-on-a-chip microfluidics approach to produce a multi-cellular endometrium in vitro, that was exposed to glucose and insulin concentrations associated with maternal metabolic stressors. Following isolation of endometrial cells (epithelial and stromal) from the uteri of non-pregnant cows in the early-luteal phase (Day 4-7 approximately), epithelial cells were seeded in the upper chamber of the device (4-6 104 cells/mL) and stromal cells seeded in the lower chamber (1.5-2 104 cells/mL). Three different concentration of glucose (0.5, 5.0 or 50 mM) or insulin (Vehicle, 1 or 10 ng/mL) were performed in the endometrial cells at a flow rate of 1µL/min for 72 hr to mimic the rate of secretion in vivo. Quantitative differences in the transcriptomic response of the cells and the secreted proteome of in vitro-derived uterine luminal fluid (ULF) were determined by RNA-sequencing and Tandem Mass Tagging Mass Spectrometry (TMT), respectively. Changes in maternal glucose altered 21 and 191 protein-coding genes in epithelial and stromal cells, respectively (p<0.05), with a dose-dependent quantitative change in the protein secretome (1 and 23 proteins in epithelial and stromal cells, respectively). Altering insulin concentrations resulted in limited transcriptional changes including transcripts for insulin-like binding proteins that were cell specific (5, 12, and 20) but altered the quantitative secretion of 196 proteins including those involved in extracellular matrix-receptor interaction and proteoglycan signaling in cancer. Collectively, these findings highlight one potential mechanism by which changes to maternal glucose and insulin associated with metabolic stress may alter uterine function.
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| Overall design |
Two different concentrations of glucose (5 or 50 mM) and three of insulin (Vehicle, 1 or 10 ng/mL) were assessed on two sets of endometrial cells (epithelial or stromal) in three biological replicates, totalizing 30 single-end fastq files (12 glucose experiment + 18 insulin experiment)
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| Contributor(s) |
DeBem T, Tinning H, Vasconcelos E, Wang D, Forde N |
| Citation(s) |
33693651 |
| Submission date |
Feb 19, 2021 |
| Last update date |
May 24, 2021 |
| Contact name |
Julie Aspden |
| E-mail(s) |
j.aspden@leeds.ac.uk
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| Organization name |
University of Leeds
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| Department |
SMCB - FBS
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| Lab |
Aspden's Lab
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| Street address |
Clarendon Way
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| City |
Leeds |
| State/province |
West Yorkshire |
| ZIP/Postal code |
LS2 9JT |
| Country |
United Kingdom |
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| Platforms (1) |
| GPL23055 |
Illumina NextSeq 500 (Bos taurus) |
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| Samples (30)
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| Relations |
| BioProject |
PRJNA702980 |
| SRA |
SRP307116 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE167086_Epi50_vs_Str50.DESeq2.format.txt.gz |
859.2 Kb |
(ftp)(http) |
TXT |
| GSE167086_Epi5_vs_Epi50.DESeq2.format.txt.gz |
741.7 Kb |
(ftp)(http) |
TXT |
| GSE167086_Epi5_vs_Str5.DESeq2.format.txt.gz |
867.5 Kb |
(ftp)(http) |
TXT |
| GSE167086_Epi_10_vs_Str_10.DESeq2.format.txt.gz |
842.6 Kb |
(ftp)(http) |
TXT |
| GSE167086_Epi_1_vs_Epi_10.DESeq2.format.txt.gz |
706.4 Kb |
(ftp)(http) |
TXT |
| GSE167086_Epi_1_vs_Str_1.DESeq2.format.txt.gz |
848.0 Kb |
(ftp)(http) |
TXT |
| GSE167086_Epi_V_vs_Epi_1.DESeq2.format.txt.gz |
709.3 Kb |
(ftp)(http) |
TXT |
| GSE167086_Epi_V_vs_Epi_10.DESeq2.format.txt.gz |
710.0 Kb |
(ftp)(http) |
TXT |
| GSE167086_Epi_V_vs_Str_V.DESeq2.format.txt.gz |
847.6 Kb |
(ftp)(http) |
TXT |
| GSE167086_Str5_vs_Str50.DESeq2.format.txt.gz |
752.1 Kb |
(ftp)(http) |
TXT |
| GSE167086_Str_1_vs_Str_10.DESeq2.format.txt.gz |
698.8 Kb |
(ftp)(http) |
TXT |
| GSE167086_Str_V_vs_Str_1.DESeq2.format.txt.gz |
696.9 Kb |
(ftp)(http) |
TXT |
| GSE167086_Str_V_vs_Str_10.DESeq2.format.txt.gz |
702.3 Kb |
(ftp)(http) |
TXT |
| GSE167086_glucose_gene_counts.txt.gz |
446.9 Kb |
(ftp)(http) |
TXT |
| GSE167086_insulin_gene_counts.txt.gz |
569.0 Kb |
(ftp)(http) |
TXT |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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