RNA sequencing analysis of FGF2-responsive transcriptome in skin fibroblasts

Abstract

Background Fibroblast growth factor 2 (FGF2) is a highly pleiotropic cytokine with antifibrotic activity in wound healing. During the process of wound healing and fibrosis, fibroblasts are the key players. Although accumulating evidence has suggested the antagonistic effects of FGF2 in the activation process of fibroblasts, the mechanisms by which FGF2 hinders the fibroblast activation remains incompletely understood. This study aimed to identify the key genes and their regulatory networks in skin fibroblasts treated with FGF2. Methods RNA-seq was performed to identify the differentially expressed mRNA (DEGs) and lncRNA between FGF2-treated fibroblasts and control. DEGs were analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Furthermore, the networks between mRNAs and lncRNAs were constructed by Pearson correlation analysis and the networkanalyst website. Finally, hub genes were validated by real time-PCR. Results Between FGF2-treated fibroblasts and control fibroblasts, a total of 1475 DEGs was obtained. These DEGs were mainly enriched in functions such as the ECM organization, cell adhesion, and cell migration. They were mainly involved in ECM-receptor interaction, PI3K-Akt signaling, and the Hippo pathway. The hub DEGs included COL3A1, COL4A1, LOX, PDGFA, TGFBI, and ITGA10. Subsequent real-time PCR, as well as bioinformatics analysis, consistently demonstrated that the expression of ITGA10 was significantly upregulated while the other five DEGs (COL3A1, COL4A1, LOX, PDGFA, TGFBI) were downregulated in FGF2-treated fibroblasts. Meanwhile, 213 differentially expressed lncRNAs were identified and three key lncRNAs (HOXA-AS2, H19, and SNHG8) were highlighted in FGF2-treated fibroblasts. Conclusion The current study comprehensively analyzed the FGF2-responsive transcriptional profile and provided candidate mechanisms that may account for FGF2-mediated wound healing.

FGF-2 enhances fibrogenetic changes in TGF-β2 treated human conjunctival fibroblasts

“…In addition, the underlying mechanisms for causing some additive effect by FGF-2 and TGF-β2 as observed in the biophysical analyses of the above 2D and 3D HconF cells remain to be elucidated despite our efforts to closely examine the several candidate molecules. Therefore, to better understand the results related to the role of FGF-2 on TGF-β2-induced fibrogenetic changes of 2D and 3D cultured HconF cells, further investigations, including an RNA-Seq experiment will be needed to elucidate possible mechanisms linked with several other complex signaling networks in order to confirm our present observations, since quite recently, in an RNA-Seq study, the FGF-2-responsive transcriptional profile was comprehensively analyzed and the findings revealed potential candidates for mechanisms in terms of FGF-2-mediated wound healing using skin fibroblasts 56 .…”

Fibroblast growth factor-2 bound to specific dermal fibroblast-derived extracellular vesicles is protected from degradation

“…File 2). Of note, among these, the most activated genes in our study ESM-1 and MMP-1 are also found in other studies reporting FGF2-induced genes 32,33 . Activation of three of those four genes by FGF2-EVs was confirmed by RT-qPCR in DFs (Fig.…”

mRNA sequencing reveals the distinct gene expression and biological functions in cardiac fibroblasts regulated by recombinant fibroblast growth factor 2

“…RNA sequencing technique had been utilized to investigate the effects of FGF2 on mRNA transcription in fibroblasts derived from the skin and lung ( Fortier et al, 2021 ; Kashpur et al, 2013 ; Wu et al, 2021a ). As described, the expression of cytoskeleton genes and ECM organization genes, e.g ., Acta2 , Myh10 , Eln , Col1a1 , Col1a2 and Postn , was significantly decreased, which is consistent with our findings ( Fortier et al, 2021 ; Kashpur et al, 2013 ; Wu et al, 2021a ). Nevertheless, fibroblasts isolated from different tissues exhibit certain heterogeneity, which may result in the diversity of gene expression regulated by FGF2.…”