Vocal Fold Fibroblasts in Reinke’s Edema Show Alterations Involved in Extracellular Matrix Production, Cytokine Response and Cell Cycle Control

Abstract

The voice disorder Reinke’s edema (RE) is a smoking- and voice-abuse associated benign lesion of the vocal folds, defined by an edema of the Reinke’s space, accompanied by pathological microvasculature changes and immune cell infiltration. Vocal fold fibroblasts (VFF) are the main cell type of the lamina propria and play a key role in the disease progression. Current therapy is restricted to symptomatic treatment. Hence, there is an urgent need for a better understanding of the molecular causes of the disease. In the present study, we investigated differential expression profiles of RE and control VFF by means of RNA sequencing. In addition, fast gene set enrichment analysis (FGSEA) was performed in order to obtain involved biological processes, mRNA and protein levels of targets of interest were further evaluated. We identified 74 differentially regulated genes in total, 19 of which were upregulated and 55 downregulated. Differential expression analysis and FGSEA revealed upregulated genes and pathways involved in extracellular matrix (ECM) remodeling, inflammation and fibrosis. Downregulated genes and pathways were involved in ECM degradation, cell cycle control and proliferation. The current study addressed for the first time a direct comparison of VFF from RE to control and evaluated immediate functional consequences.

Effects of Electronic (E)‐cigarette Vapor and Cigarette Smoke in Cultured Vocal Fold Fibroblasts

“…These cells were targeted as they are the main cell type of the lamina propria and implicated in the development of CSinduced vocal fold disease. 14,16 In addition, from a practical standpoint, at the time, this study was conducted, immortalized hVFFs were readily available for experimental use. However, we acknowledge that epithelial cells are the first cell type in the vocal folds to encounter EV and CS and critical modulators of vocal fold disease development.…”

“…Here, we focused on the effects of EVE and CSE on hVFFs. These cells were targeted as they are the main cell type of the lamina propria and implicated in the development of CS‐induced vocal fold disease 14,16 . In addition, from a practical standpoint, at the time, this study was conducted, immortalized hVFFs were readily available for experimental use.…”

“…13 VFF are implicated in the development of CS-induced vocal fold diseases, including Reinke's edema. [14][15][16] VFF are also among the first cells exposed to EV. No studies to date have investigated whether e-cigs alter the function of these cells; consequently, more work is needed to elucidate how VFF may contribute to e-cig-induced changes in vocal fold health and disease development.…”

“…Vocal fold fibroblasts (VFF), the primary cell type in the lamina propria, are important mediators of inflammation and necessary for maintenance of the extracellular matrix (ECM) of the lamina propria 13 . VFF are implicated in the development of CS‐induced vocal fold diseases, including Reinke's edema 14–16 . VFF are also among the first cells exposed to EV.…”

A universal co‐expression gene network and prognostic model for hepatic–biliary–pancreatic cancers identified by integrative analyses

“…Gene Set Enrichment Analysis (GSEA) against hallmark gene sets in MsigDb [25] was performed using genes ranked by log2-transformed fold change (log2FC) with the FGSEA package [26] in R. To evaluate the potential functions of uniformly upregulated or downregulated genes of the cancers, Gene Ontology (GO; http://geneontology.org) and Kyoto Encyclopedia of Genes and Genomes (KEGG; https://www.genome.jp/kegg) pathway enrichment analysis was performed using the CLUSTERPROFILER package in R [27]. The significant threshold value was set as P < 0.05.…”