Multi-omics analysis reveals regulators of the response to PDGF-BB treatment in pulmonary artery smooth muscle cells

Abstract

BackgroundPulmonary arterial hypertension (PAH) is a lethal disease with pronounced narrowing of pulmonary vessels due to abnormal cell proliferation. The platelet-derived growth factor BB (PDGF-BB) is well known as a potent mitogen for smooth muscle cell proliferation. To better understand how this growth factor regulates pulmonary arterial smooth muscle cells (PASMCs) proliferation, we sought to characterize the response to PDGF-BB stimulation at system-wide levels, including the transcriptome and proteome.ResultsIn this study, we identified 1611 mRNAs (transcriptome), 207 proteins (proteome) differentially expressed in response to PDGF-BB stimulation in PASMCs based on RNA-sequencing and isobaric tags for relative and absolute quantification (iTRAQ) assay. Transcription factor (TF)-target network analysis revealed that PDGF-BB regulated gene expression potentially via TFs including HIF1A, JUN, EST1, ETS1, SMAD1, FOS, SP1, STAT1, LEF1 and CEBPB. Among them, SMAD1-involved BMPR2/SMADs axis plays a significant role in PAH development. Interestingly, we observed that the expression of BMPR2 was decreased in both mRNA and protein level in response to PDGF-BB. Further study revealed that BMPR2 is the direct target of miR-376b that is up-regulated upon PDGF-BB treatment. Finally, EdU incorporation assay showed that miR-376b promoted proliferation of PASMCs.ConclusionThis integrated analysis of PDGF-BB-regulated transcriptome and proteome was performed for the first time in normal PASMCs, which revealed a crosstalk between PDGF signaling and BMPR2/SMADs axis. Further study demonstrated that PDGF-BB-induced miR-376b upregulation mediated the downregulation of BMPR2, which led to expression change of its downstream targets and promoted proliferation of PASMCs.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3122-3) contains supplementary material, which is available to authorized users.

SphK1/S1P Mediates PDGF-Induced Pulmonary Arterial Smooth Muscle Cell Proliferation via miR-21/BMPRII/Id1 Signaling Pathway

“…Downregulation of BMPRII protein reduces the expression of Id1 gene, which further promotes PASMC proliferation [37,38]. Recent study has demonstrated that PDGF inhibits BMPRII/ Id1 signaling and thus promotes PASMC proliferation [39]. It has been shown that miR-21 reduces BMPRII protein by directly targeting BMPRII mRNA, promotes PASMC proliferation and further regulates pulmonary vascular remodeling [5,30,40].…”

MiR-339 inhibits proliferation of pulmonary artery smooth muscle cell by targeting FGF signaling

“…MiR‐339 is one of the differentially expressed miRNAs reported in our previous work (Chen et al. ). In this study, miR‐339 was found to be downregulated not only by PDGF‐BB, but also by many other cytokines and growth factors, including FGF2, ET‐1, ANG II, and so forth.…”

“…Recently, we have identified 13 miRNAs that were differentially expressed in rat PASMC (RPASMC) in response to PDGF‐BB stimulation (Chen et al. ). The expression level of these miRNAs were measured in different organs of the rat and one miRNA, miR‐339, was found to be relatively highly expressed in the arteries, compared to most other organs (Fig.…”

“…), and miR‐376b expression was increased by PDGF‐BB, a miRNA that increased proliferation of PASMC via targeting BMPR2 (Chen et al. ). MiR‐339 is one of the differentially expressed miRNAs reported in our previous work (Chen et al.…”

“…For example, our previous report indicated a cross‐talk between PDGF and BMP signaling mediated by miR‐376b (Chen et al. ). In this work, we also discovered a cross‐talk between FGF signaling and other factors, including PDGF‐BB, mediated by miR‐339 via targeting FRS2.…”