RNA-sequencing demonstrates transcriptional differences between human vocal fold fibroblasts and myofibroblasts. RNA-sequencing demonstrates transcriptional differences between human vocal fold fibroblasts and myofibroblasts

Background: Differentiation of fibroblasts into myofibroblasts is necessary for wound healing, but excessive myofibroblast presence and persistence can result in scarring. Treatment for scarring is limited largely due to a lack of comprehensive understanding of how fibroblasts and myofibroblasts differ at the transcript level. The purpose of the study was to comprehensively characterize transcriptional profiles of injured fibroblasts relative to normal fibroblasts, utilizing fibroblasts from the vocal fold as a model for other areas of the body. Results: we identified differentially expressed genes between groups of normal fibroblasts, scarred fibroblasts, and fibroblasts treated with transforming growth factor-beta 1 (TGF-β1), which represented an induced-scar phenotype. Principal component analysis revealed clustering of normal fibroblasts separate from the clustering of fibroblasts treated with TGF-β1; scarred fibroblasts were more similar to normal fibroblasts than fibroblasts treated with TGF-β1. Enrichment analyses revealed pathways related to cell signaling, receptor-ligand activity, and regulation of cell functions in scarred fibroblasts, pathways related to cell adhesion in normal fibroblasts, and pathways related to ECM binding in fibroblasts treated with TGF-β1. Although transcriptomic profiles between scarred fibroblasts and fibroblasts treated with TGF-β1 were relatively dissimilar, the most highly co-expressed genes were enriched in pathways related to actin cytoskeleton binding. Conclusions: Transcriptomics of normal fibroblasts differ from myofibroblasts, including from those retrieved from scar and those treated with TGF-β1. Despite large differences in transcriptomics between tVFF and sVFF, tVFF serve as a useful in vitro model of myofibroblasts and highlight key similarities to myofibroblasts extracted from scar pathology, as well as expected differences related to normal fibroblasts from healthy VF. Overall design: We collected normal fibroblasts, scarred fibroblasts, and fibroblasts treated with transforming growth factor-beta 1 (TGF-β1), which represented an induced-scar phenotype. We then performed gene expression profiling analysis, and analyzed their differences using gene expression analysis methods. *************************************************************** Submitter states that missing raw files are due to file corruption. ***************************************************************

背景：成纤维细胞向肌成纤维细胞的分化是伤口愈合的必要环节，但肌成纤维细胞过度蓄积及持续存留可引发瘢痕形成。当前瘢痕治疗手段极为有限，核心原因在于对成纤维细胞与肌成纤维细胞在转录层面的差异缺乏全面认知。本研究以声带成纤维细胞作为人体其他部位组织的研究模型，旨在全面表征损伤性成纤维细胞相较于正常成纤维细胞的转录组谱。
结果：我们筛选得到正常成纤维细胞、瘢痕来源成纤维细胞，以及经转化生长因子-β1（transforming growth factor-beta 1, TGF-β1）处理的成纤维细胞（该处理可诱导出瘢痕表型）这三组样本间的差异表达基因。主成分分析结果显示，正常成纤维细胞的聚类结果与经TGF-β1处理的成纤维细胞显著分离；相较于经TGF-β1处理的成纤维细胞，瘢痕来源成纤维细胞与正常成纤维细胞的相似性更高。富集分析结果表明：瘢痕来源成纤维细胞富集了与细胞信号转导、受体-配体活性及细胞功能调控相关的通路；正常成纤维细胞富集了与细胞黏附相关的通路；而经TGF-β1处理的成纤维细胞则富集了与细胞外基质（extracellular matrix, ECM）结合相关的通路。尽管瘢痕来源成纤维细胞与经TGF-β1处理的成纤维细胞的转录组谱存在相对显著的差异，但二者共表达程度最高的基因富集于肌动蛋白细胞骨架结合相关通路。
结论：正常成纤维细胞的转录组谱与肌成纤维细胞存在显著差异，后者既包括瘢痕来源的肌成纤维细胞，也包括经TGF-β1诱导的肌成纤维细胞。尽管经TGF-β1处理的声带成纤维细胞（tVFF）与瘢痕来源声带成纤维细胞（sVFF）之间的转录组差异显著，但tVFF仍可作为一种实用的肌成纤维细胞体外模型，其既与瘢痕病理来源的肌成纤维细胞存在关键共性，同时也与健康声带来源的正常成纤维细胞存在预期的差异。
整体实验设计：我们收集了正常成纤维细胞、瘢痕来源成纤维细胞，以及经转化生长因子-β1（TGF-β1）处理的成纤维细胞（该处理可诱导瘢痕表型），随后开展基因表达谱分析，并通过基因表达分析方法对各组间的表达差异进行了探究。
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