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| Status |
Public on Mar 01, 2021 |
| Title |
ASCL2 reciprocally controls key trophoblast lineage decisions during hemochorial placenta development |
| Organisms |
Homo sapiens; Rattus norvegicus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Invasive trophoblast cells are critical to spiral artery remodeling in hemochorial placentation. Insufficient trophoblast invasion and vascular remodeling can lead to pregnancy disorders including preeclampsia, preterm birth, and intrauterine growth restriction. Previous studies in the mouse identified achaete-scute homolog 2 (ASCL2) as essential to extraembryonic development. We hypothesized that ASCL2 is a critical and conserved regulator of invasive trophoblast lineage development. In contrast to the mouse, the rat possesses deep intrauterine trophoblast cell invasion and spiral artery remodeling similar to human placentation. In this report, we investigated invasive/extravillous trophoblast (EVT) cell differentiation using human trophoblast stem (TS) cells and a loss-of-function mutant Ascl2 rat model. ASCL2 transcripts are expressed in the EVT column and junctional zone, which represent tissue sources of invasive trophoblast progenitor cells within human and rat placentation sites, respectively. Differentiation of human TS cells into EVT cells resulted in significant upregulation of ASCL2 and several other transcripts indicative of EVT cell differentiation. Disruption of ASCL2 impaired EVT cell differentiation as indicated by cell morphology and transcript profiles. RNA sequencing analysis of ASCL2-deficient trophoblast cells identified both downregulation of EVT cell-associated transcripts and upregulation of syncytiotrophoblast-associated transcripts, indicative of dual activating and repressing functions. ASCL2 deficiency in the rat impacted placental morphogenesis resulting in junctional zone dysgenesis and failed intrauterine trophoblast cell invasion. ASCL2 acts as a critical and conserved regulator of invasive trophoblast cell lineage development and a species-specific modulator of the syncytiotrophoblast lineage.
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| Overall design |
1) ASCL2 was knocked down in human trophoblast stem (TS) cells using lentiviral delivery of shRNA. Following selection by puromycin, human TS cells were differentiated for eight days (Okae et al.) into extravillous trophoblast (EVT). On day eight of differentiation, cells treated with either control shRNA or ASCL2-specific shRNA were collected and RNA was isolated. 2) Rat placental disc tissues were dissected on gd12.5 from heterozygous breeding of Ascl2 mutant rats. Dissected placentation site compartments from wild type and Ascl2 null fetuses were snap-frozen in liquid nitrogen and stored at -80°C until processing for RNA isolation.
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| Contributor(s) |
Varberg KM, Iqbal K, Muto M, Simon ME, Scott RL, Kozai K, Choudhury RH, Aplin JD, Biswell R, Gibson M, Okae H, Arima T, Vivian JL, Grundberg E, Soares MJ |
| Citation(s) |
33649217 |
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| Submission date |
Jul 13, 2020 |
| Last update date |
Mar 03, 2021 |
| Contact name |
Kaela Margaret Varberg |
| E-mail(s) |
kaelasevera@gmail.com, kmvarberg@cmh.edu
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| Phone |
7852181354
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| Organization name |
Children's Mercy
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| Department |
Pediatrics
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| Lab |
Varberg Lab
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| Street address |
2401 Gillham Road; CMRI 7902.19
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| City |
Kansas City |
| State/province |
MO |
| ZIP/Postal code |
64108 |
| Country |
USA |
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| Platforms (2) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
| GPL25947 |
Illumina NovaSeq 6000 (Rattus norvegicus) |
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| Samples (17)
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| Relations |
| BioProject |
PRJNA645927 |
| SRA |
SRP271644 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE154350_ASCL2_hTS_processed_data.xlsx |
18.3 Mb |
(ftp)(http) |
XLSX |
| GSE154350_ASCL2_rat_processed_data_kv.xlsx |
5.2 Mb |
(ftp)(http) |
XLSX |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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