Mullin Nathaniel K, Mallipeddi Nikhil V, Hamburg-Shields Emily, Ibarra Beatriz, Khalil Ahmad M, Atit Radhika P
Department of Biology, Case Western Reserve University, Cleveland, OH, United States.
Department of Genetics and Genome Sciences, School of Medicine, Case Western Reserve University, Cleveland, OH, United States.
Front Genet. 2017 Nov 21;8:183. doi: 10.3389/fgene.2017.00183. eCollection 2017.
Wnt/β-catenin signaling is required for embryonic dermal fibroblast cell fate, and dysregulation of this pathway is sufficient to promote fibrosis in adult tissue. The downstream modulators of Wnt/β-catenin signaling required for controlling cell fate and dermal fibrosis remain poorly understood. The discovery of regulatory long non-coding RNAs (lncRNAs) and their pivotal roles as key modulators of gene expression downstream of signaling cascades in various contexts prompted us to investigate their roles in Wnt/β-catenin signaling. Here, we have identified lncRNAs and protein-coding RNAs that are induced by β-catenin activity in mouse dermal fibroblasts using next generation RNA-sequencing. The differentially expressed protein-coding mRNAs are enriched for extracellular matrix proteins, glycoproteins, and cell adhesion, and many are also dysregulated in human fibrotic tissues. We identified 111 lncRNAs that are differentially expressed in response to activation of Wnt/β-catenin signaling. To further characterize the role of mouse lncRNAs in this pathway, we validated two novel Wnt signaling- Induced Non-Coding RNA (Wincr) transcripts referred to as and . These two lncRNAs are highly expressed in mouse embryonic skin and perinatal dermal fibroblasts. Furthermore, we found that expression levels in perinatal dermal fibroblasts affects the expression of key markers of fibrosis (e.g., and ), enhances collagen contraction, and attenuates collective cell migration. Our results show that β-catenin signaling-responsive lncRNAs may modulate dermal fibroblast behavior and collagen accumulation in dermal fibrosis, providing new mechanistic insights and nodes for therapeutic intervention.
Wnt/β-连环蛋白信号通路对于胚胎真皮成纤维细胞的细胞命运是必需的，并且该信号通路的失调足以促进成体组织中的纤维化。对于控制细胞命运和真皮纤维化所需的Wnt/β-连环蛋白信号通路的下游调节因子仍知之甚少。在各种情况下，调控性长链非编码RNA（lncRNA）的发现及其作为信号级联下游基因表达关键调节因子的关键作用促使我们研究它们在Wnt/β-连环蛋白信号通路中的作用。在此，我们使用下一代RNA测序鉴定了小鼠真皮成纤维细胞中由β-连环蛋白活性诱导的lncRNA和蛋白质编码RNA。差异表达的蛋白质编码mRNA富含细胞外基质蛋白、糖蛋白和细胞粘附相关蛋白，并且许多在人类纤维化组织中也失调。我们鉴定出111种lncRNA在Wnt/β-连环蛋白信号通路激活后差异表达。为了进一步表征小鼠lncRNA在该信号通路中的作用，我们验证了两种新型的Wnt信号诱导非编码RNA（Wincr）转录本，分别称为 和 。这两种lncRNA在小鼠胚胎皮肤和围产期真皮成纤维细胞中高度表达。此外，我们发现围产期真皮成纤维细胞中的 表达水平会影响纤维化关键标志物（如 和 ）的表达，增强胶原收缩，并减弱集体细胞迁移。我们的结果表明，β-连环蛋白信号响应性lncRNA可能调节真皮纤维化中真皮成纤维细胞的行为和胶原积累，为治疗干预提供了新的机制见解和靶点。
