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GEO help: Mouse over screen elements for information. |
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| Status |
Public on Jan 12, 2019 |
| Title |
Next-generation sequencing of murine trophoblast-derived and pregnancy-associated exosome-enriched extracellular vesicle microRNAs |
| Organism |
Mus musculus |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
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| Summary |
The role of extracellular vesicles (EVs), specifically exosomes, in intercellular communication likely plays a key role in placental orchestration of pregnancy and maternal immune sensing of the fetus. While murine models are powerful tools to study pregnancy and maternal-fetal immune interactions, in contrast to human placental exosomes, the content of murine placental and pregnancy exosomes remains largely understudied. Using a recently developed in vitro culture technique, murine trophoblast stem cells derived from B6 mice were differentiated into syncytial-like cells. EVs from the conditioned media, as well as from pregnant and non-pregnant sera, were enriched for exosomes. The RNA composition of these murine trophoblast-derived and pregnancy-associated exosome-enriched-EVs (ExoE-EVs) was determined using RNA-sequencing analysis and expression levels confirmed by qRT-PCR. Differentially abundant miRNAs were detected in syncytial differentiated ExoE-EVs, particularly from the X chromosome cluster (mmu-miR-322-3p, mmu-miR-322-5p, mmu-miR-503-5p, mmu-miR-542-3p, and mmu-miR-450a-5p). These were confirmed to be increased in pregnant mouse sera ExoE-EVs by qRT-PCR analysis. Interestingly, fifteen miRNAs were only present within the pregnancy-derived ExoE-EVs compared to non-pregnant controls. Mmu-miR-292-3p and mmu-miR-183-5p were noted to be some of the most abundant miRNAs in syncytial ExoE-EVs and were also present at higher levels in pregnant versus non-pregnant sera ExoE-EVs. The bioinformatics tool, MultiMir, was employed to query publicly available databases of predicted miRNA-target interactions. This analysis reveals that the X-chromosome miRNAs are predicted to target ubiquitin-mediated proteolysis and intracellular signaling pathways. Knowing the cargo of placental and pregnancy-specific ExoE-EVs as well as the predicted biological targets informs studies using murine models to examine not only maternal-fetal immune interactions but also the physiologic consequences of placental-maternal communication.
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| Overall design |
Examination of EVs isolated from 1) conditioned media from in vitro mouse trophoblast cells cultures maintained as "STEM" using Cell Start and FGF4 (n=3), 2) conditioned media from in vitro mouse trophoblast cultures that have undergone syncytial-like differentiation (n=3), 3) non-pregnant B6 mouse sera (n=3); 4) pregnant B6 mouse sera (gestational day 14.5, n=3). EVs were isolated using ExoQuick-TC (1-2) or ExoQuick (3-4).
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| Contributor(s) |
Stefanski AL, Martinez N, Peterson LK, Callahan TJ, Treacy E, Luck M, Friend SF, Hermesch A, Maltepe E, Phang T, Dragone LL, Winn VD |
| Citation(s) |
30730971 |
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| Submission date |
Jan 11, 2019 |
| Last update date |
Mar 06, 2019 |
| Contact name |
Virginia D. Winn |
| E-mail(s) |
[email protected]
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| Organization name |
Stanford University
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| Department |
OB-GYN/Reproductive, Perinatal and Stem Cell Biology Research
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| Street address |
300 Pasteur Drive, Boswell A364
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| City |
Stanford |
| State/province |
CA |
| ZIP/Postal code |
94305 |
| Country |
USA |
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| Platforms (1) |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA514778 |
| SRA |
SRP178560 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE124995_Abundances_Day14.5.txt.gz |
9.3 Mb |
(ftp)(http) |
TXT |
| GSE124995_Abundances_Diff.txt.gz |
10.1 Mb |
(ftp)(http) |
TXT |
| GSE124995_Abundances_NP.txt.gz |
9.1 Mb |
(ftp)(http) |
TXT |
| GSE124995_Abundances_Stem.txt.gz |
10.3 Mb |
(ftp)(http) |
TXT |
| GSE124995_NPvsDay14.5_all_with_type.1384366252915.tsv.gz |
9.9 Mb |
(ftp)(http) |
TSV |
| GSE124995_StemvsDiff_all_with_type.1384366250358.tsv.gz |
13.2 Mb |
(ftp)(http) |
TSV |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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