Transcriptome Analysis Reveals an Inhibitory Effect of Dihydrotestosterone-Treated 2D- and 3D-Cultured Dermal Papilla Cells on Hair Follicle Growth

Abstract

Dermal papillae are a target of androgen action in patients with androgenic alopecia, where androgen acts on the epidermis of hair follicles in a paracrine manner. To mimic the complexity of the dermal papilla microenvironment, a better culture model of human dermal papilla cells (DPCs) is needed. Therefore, we evaluated the inhibitory effect of dihydrotestosterone (DHT)-treated two-dimensional (2D)- and 3D-cultured DPCs on hair follicle growth. 2D- and 3D-cultured DPC proliferation was inhibited after co-culturing with outer root sheath (ORS) cells under DHT treatment. Moreover, gene expression levels of β-catenin and neural cell adhesion molecules were significantly decreased and those of cleaved caspase-3 significantly increased in 2D- and 3D-cultured DPCs with increasing DHT concentrations. ORS cell proliferation also significantly increased after co-culturing in the control-3D model compared with the control-2D model. Ki67 downregulation and cleaved caspase-3 upregulation in DHT-treated 2D and 3D groups significantly inhibited ORS cell proliferation. Sequencing showed an increase in the expression of genes related to extracellular matrix synthesis in the 3D model group. Additionally, the top 10 hub genes were identified, and the expression of nine chemokine-related genes in DHT-treated DPCs was found to be significantly increased. We also identified the interactions between transcription factor (TF) genes and microRNAs (miRNAs) with hub genes and the TF–miRNA coregulatory network. Overall, the findings indicate that 3D-cultured DPCs are more representative of in vivo conditions than 2D-cultured DPCs and contribute to our understanding of the molecular mechanisms underlying androgen-induced alopecia.

Effects of photobiomodulation at various irradiances on normal and dihydrotestosterone‐treated human hair dermal papilla cells in vitro

“…50 μM DHT significantly decreased the cell viability of DPCs, suggesting this concentration could simulate the negative effects of DHT on DPCs. The experimental results align with previous studies that demonstrate the ability of high concentrations of DHT (10 μM) to simulate the dermal papilla microenvironment in androgen‐induced alopecia [20, 21]. Therefore, for subsequent experiments, a concentration of 50 μM DHT was selected for treating DPCs.…”

“…Overall, our results suggested that PBM had certain downregulating effects on the growth-inhibitory genes in normal DPCs but the optimal irradiance varied among PBM groups within different aspects. the dermal papilla microenvironment in androgeninduced alopecia [20,21]. Therefore, for subsequent experiments, a concentration of 50 μM DHT was selected for treating DPCs.…”

Direct Reprograming of Mouse Fibroblasts into Dermal Papilla Cells via Small Molecules

“…KEGG analysis of the up-regulated DEGs in treated vs. control group showed the top 20 enriched pathways, including platelet activation, the Rap1 signaling pathway, cytokine-cytokine receptor interaction, cell adhesion molecules (CAMs), the PPAR signaling pathway, ECM-receptor interaction, vascular smooth muscle contraction and neuroactive ligand-receptor interaction (Figure 5b). An increasing amount of research suggests an interaction between chemokines and skin appendage morphogenesis [41][42][43]. Moreover, the result of the cytokine-cytokine receptor interaction pathway was also enriched in the TTNPB-treated group.…”

“…The hub genes of at least five algorithms that overlapped were obtained, containing ten hub genes (Il6, Cxcl12, Il15, Tgfb1, Cxcl1, Il18, Ccl11, Cxcl10, Cxcl13 and Cxcl5) (Figure 5c). The expression of An increasing amount of research suggests an interaction between chemokines and skin appendage morphogenesis [41][42][43]. Moreover, the result of the cytokine-cytokine receptor interaction pathway was also enriched in the TTNPB-treated group.…”

Editorial: Hair Follicle Stem Cell Regeneration in Aging

“…As two-dimensional (2D) cultured cells quickly lose their regenerative ability, most researches turned to modulate hair regeneration under 3D culture conditions. Zhang et al compared the gene expression in 2D and 3D cultured DP cells treated with dihydrotestosterone, which can inhibit proliferation but induce apoptosis in DP cells co-cultured with ORS cells ( Zhang Y. et al, 2021 ). Without dihydrotestosterone treatment, the ORS cells showed significant proliferation when co-cultured with DP cells.…”