Dataset Information

The long non-coding RNA DAGAR is regulated by m6A modification and both pathways are important for smooth muscle cell plasticity

ABSTRACT: Vascular smooth muscle cells (SMCs) change between a contractile-differentiated and a proliferative-dedifferentiated phenotype in response to environmental cues. Long non-coding RNAs (lncRNAs) and N6-methyladenosine (m6A) modification regulate cell fate decision. Here we used primary human pulmonary artery SMCs (hPASMCs) and assessed the expression of lncRNAs during SMCs phenotypic modulation. A lncRNA, which we refer to as Differentiation And Growth Arrest-related lncRNA (DAGAR) was increased during differentiation and we demonstrate that it is required for this process. DAGAR was m6A-modified and regulated by the m6A reader YTHDF2 in SMCs and MRC5 cells. A marked downregulation of YTHDF1-3 proteins during both SMC differentiation and MRC5 quiescence was found, consistent with the increase of DAGAR. Remarkably, YTHDF2 immunoprecipitation followed by RNA deep sequencing (RIP-Seq) displayed an enrichment of key SMC-associated transcripts, including smooth muscle myosin heavy chain (MYH11) and members of the TGF, PDGF and VEGF pathways. Knockdown of YTHDF2 induced DAGAR and SMC marker gene expression. We conclude that the lncRNA DAGAR and YTHDF2 contribute to the regulation of SMC plasticity and differentiation programs.

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ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Primary Cell

SUBMITTER: Astrid Bruckmann

LAB HEAD: Astrid Bruckmann

PROVIDER: PXD029550 | Pride | 2025-05-06

REPOSITORIES: Pride

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Publications

Differentiation and Growth-Arrest-Related lncRNA (<i>DAGAR</i>): Initial Characterization in Human Smooth Muscle and Fibroblast Cells.

de la Cruz-Thea Benjamin B Natali Lautaro L Ho-Xuan Hung H Bruckmann Astrid A Coll-Bonfill Núria N Strieder Nicholas N Peinado Víctor I VI Meister Gunter G Musri Melina M MM

International journal of molecular sciences 20240831 17

Vascular smooth muscle cells (SMCs) can transition between a quiescent contractile or "differentiated" phenotype and a "proliferative-dedifferentiated" phenotype in response to environmental cues, similar to what in occurs in the wound healing process observed in fibroblasts. When dysregulated, these processes contribute to the development of various lung and cardiovascular diseases such as Chronic Obstructive Pulmonary Disease (COPD). Long non-coding RNAs (lncRNAs) have emerged as key modulator ...[more]

PMID: 39273443

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Project description:Tissue Factor Pathway Inhibitor-2 is Induced by Fluid Shear Stress in Vascular Smooth Muscle Cells and Affects Cell Proliferation and Survival Introduction: Vascular smooth muscle cells (SMCs) are exposed to fluid shear stress (FSS) after interventional procedures such as balloon-angioplasty. Whereas the effects of hemodynamic forces on endothelial cells are explored in detail, the influence of FSS on smooth muscle cell function is poorly characterized. Here, we investigated the effect of FSS on SMC gene expression and function. Methods: Laminar FSS of arterial level (14 dynes/cm2) was applied to SMC cultures for 24 h in a parallel-plate flow chamber. The effect of FSS on gene expression was first screened with microarray technology and the results further verified by real time (RT) PCR and immunoblotting. Protein expression was also studied in the rat carotid artery after balloon-injury and DNA synthesis and apoptosis was examined in SMCs in vitro. Results: Microarrays identified tissue-pathway inhibitor-2 (TFPI-2) as the most differentially expressed gene by FSS in cultured SMCs. The regulatory effect of FSS on the expression of TFPI-2 was confirmed by RT-PCR and immunobloting demonstrating a more than 400-fold increase in TFPI-2 expression in SMCs exposed to FSS compared to static controls and a consistent upregulation at the protein level. Functionally, SMC proliferation was decreased by FSS and recombinant TFPI-2 was found to inhibit SMC proliferation and induce SMC apoptosis as indicated by activation of caspase-3. In vivo, TFPI-2 expression was found to be up-regulated 5, 10 and 20 h after rat carotid balloon-injury and immunohistochemistry demonstrated TFPI-2 protein in luminal SMCs exposed to FSS in rat carotid intimal hyperplasia 10 days after balloon-injury. Conclusion: FSS strongly influence gene expression in cultured SMCs and induce TFPI-2 expression, which is also expressed after rat carotid balloon injury in luminal SMCs exposed to FSS. Functionally, TFPI-2 may play an important role in vessel wall repair by regulating SMC proliferation and survival. Further studies are needed to elucidate the mechanisms by which TFPI-2 control SMC function. 3 x 2 samples from a paired fluid shear stress experiment on smooth muscle cells.

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