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Updated: Dec 7, 2025
Molecular Analysis of Endothelial-mesenchymal Transition Induced by Transforming Growth Factor-β Signaling
Published on: August 3, 2018
Impact of Sustained Transforming Growth Factor-β Receptor Inhibition on Chromatin Accessibility and Gene Expression
Raffaella Lucciola1,2,3, Pavle Vrljicak3,4, Shanti Gurung1
1The Ritchie Centre, Hudson Institute of Medical Research, Melbourne, VIC, Australia.
Abstract:
Endometrial mesenchymal stem cells (eMSC) drive the extraordinary regenerative capacity of the human endometrium. Clinical application of eMSC for therapeutic purposes is hampered by spontaneous differentiation and cellular senescence upon large-scale expansion in vitro. A83-01, a selective transforming growth factor-β receptor (TGFβ-R) inhibitor, promotes expansion of eMSC in culture by blocking differentiation and senescence, but the underlying mechanisms are incompletely understood. In this study, we combined RNA-seq and ATAC-seq to study the impact of sustained TGFβ-R inhibition on gene expression and chromatin architecture of eMSC. Treatment of primary eMSC with A83-01 for 5 weeks resulted in differential expression of 1,463 genes. Gene ontology analysis showed enrichment of genes implicated in cell growth whereas extracellular matrix genes and genes involved in cell fate commitment were downregulated. ATAC-seq analysis demonstrated that sustained TGFβ-R inhibition results in opening and closure of 3,555 and 2,412 chromatin loci, respectively. Motif analysis revealed marked enrichment of retinoic acid receptor (RAR) binding sites, which was paralleled by the induction of RARB, encoding retinoic acid receptor beta (RARβ). Selective RARβ inhibition attenuated proliferation and clonogenicity of A83-01 treated eMSC. Taken together, our study provides new insights into the gene networks and genome-wide chromatin changes that underpin maintenance of an undifferentiated phenotype of eMSC in prolonged culture.
Insights
Transforming growth factor-β receptor (TGFβ-R) inhibition with A83-01 maintains undifferentiated endometrial mesenchymal stem cells (eMSC) in culture. This study reveals gene expression and chromatin changes, including retinoic acid receptor beta (RARβ) induction, crucial for eMSC expansion.
Area of Science:
- Stem cell biology
- Regenerative medicine
- Epigenetics
Background:
- Endometrial mesenchymal stem cells (eMSC) possess significant regenerative potential but face challenges in clinical expansion due to differentiation and senescence.
- Understanding the molecular mechanisms regulating eMSC proliferation and differentiation is crucial for therapeutic applications.
Purpose of the Study:
- To investigate the molecular mechanisms by which sustained transforming growth factor-β receptor (TGFβ-R) inhibition using A83-01 maintains the undifferentiated state of eMSC during prolonged in vitro culture.
- To elucidate the impact of TGFβ-R inhibition on gene expression profiles and genome-wide chromatin accessibility in eMSC.
Main Methods:
- Primary eMSC were treated with A83-01 for 5 weeks.
- RNA-sequencing (RNA-seq) was performed to analyze gene expression changes.
- Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq) was used to assess chromatin architecture.
- Gene ontology and motif analyses were conducted.
Main Results:
- Sustained TGFβ-R inhibition led to differential expression of 1,463 genes, with enrichment in cell growth pathways and downregulation of extracellular matrix and cell fate commitment genes.
- ATAC-seq revealed significant alterations in chromatin accessibility, with thousands of loci opening or closing.
- Motif analysis identified an enrichment of retinoic acid receptor (RAR) binding sites, correlating with induced expression of the RAR-beta (RARB) gene.
- Selective inhibition of RARβ reversed the proliferative and clonogenic effects of A83-01 on eMSC.
Conclusions:
- Sustained TGFβ-R inhibition preserves the undifferentiated phenotype of eMSC by modulating gene networks and chromatin accessibility.
- The retinoic acid signaling pathway, particularly RARβ, plays a critical role in mediating the effects of TGFβ-R inhibition on eMSC proliferation and maintenance.
- These findings offer novel insights into strategies for expanding eMSC for therapeutic applications.
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