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

Yufan Zhang,Zhen Liu,Yong Miao,Kaitao Li,Jin Wang,Danlan Fu,Zhiqi Hu,Junfei Huang,Hailin Wang,Qian Qu,Zhexiang Fan

doi:10.3389/fcell.2021.724310

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.

Highlights

Hair follicle development and growth depends on reciprocal epithelial–mesenchymal interactions, and the starting point of the initial signals is thought to originate from the mesenchymal dermis (Dhouailly, 1973)
The results of immunofluorescence staining of dermal papilla cells (DPCs) at passage (P)2, P4, P6, and P8 showed that the androgen receptors (ARs) protein was significantly expressed in P2 and P4 compared with that in P6 and P8 (Figure 1A)
When DPCs were treated with 10 μm DHT, immunofluorescence staining revealed that the AR had been transferred from the original cytoplasm to the nucleus (Figures 1D,E)

Summary

Introduction

Hair follicle development and growth depends on reciprocal epithelial–mesenchymal interactions, and the starting point of the initial signals is thought to originate from the mesenchymal dermis (Dhouailly, 1973). The dermal papillae, a cluster of mesenchymal cells at the base of the hair follicle, are considered to play an important role in the hair cycle by regulating the growth and activity of Inhibitory Effect of DHT-Inducible DP Cells various cells in hair follicles through the secretion of diffusible proteins to the epidermis of hair follicles (Andl et al, 2002). AGA is characterized by the gradual miniaturization of hair follicles and a premature transition from anagen to catagen induced by androgens (Jahoda, 1998). DPCs are the targets of androgen action in AGA hair follicles. Androgen drives DPCs to act on themselves in a paracrine manner in hair follicle epithelial cells, resulting in the miniaturization of hair follicles and changes in the hair follicle cycle (Randall et al, 2001). Altering the expression of DPC-related genes under DHT treatment may be a key factor in androgenpotentiated balding

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Results

Conclusion