miR-140-5p in Small Extracellular Vesicles From Human Papilla Cells Stimulates Hair Growth by Promoting Proliferation of Outer Root Sheath and Hair Matrix Cells

Abstract

The application of dermal papilla cells to hair follicle (HF) regeneration has attracted a great deal of attention. However, cultured dermal papilla cells (DPCs) tend to lose their capacity to induce hair growth during passage, restricting their usefulness. Accumulating evidence indicates that DPCs regulate HF growth mainly through their unique paracrine properties, raising the possibility of therapies based on extracellular vesicles (EVs). In this study, we explored the effects of EVs from high- and low-passage human scalp follicle dermal papilla cells (DP-EVs) on activation of hair growth, and investigated the underlying mechanism. DP-EVs were isolated by ultracentrifugation and cultured with human scalp follicles, hair matrix cells (MxCs), and outer root sheath cells (ORSCs), and we found low-passage DP-EVs accelerated HF elongation and cell proliferation activation. High-throughput miRNA sequencing and bioinformatics analysis identified 100 miRNAs that were differentially expressed between low- (P3) and high- (P8) passage DP-EVs. GO and KEGG pathway analysis of 1803 overlapping target genes revealed significant enrichment in the BMP/TGF-β signaling pathways. BMP2 was identified as a hub of the overlapping genes. miR-140-5p, which was highly enriched in low-passage DP-EVs, was identified as a potential regulator of BMP2. Direct repression of BMP2 by miR-140-5p was confirmed by dual-luciferase reporter assay. Moreover, overexpression and inhibition of miR-140-5p in DP-EVs suppressed and increased expression of BMP signaling components, respectively, indicating that this miRNA plays a critical role in hair growth and cell proliferation. DP-EVs transport miR-140-5p from DPCs to epithelial cells, where it downregulates BMP2. Therefore, DPC-derived vesicular miR-140-5p represents a therapeutic target for alopecia.

Application of Cell-Derived Extracellular Vesicles and Engineered Nanovesicles for Hair Growth: From Mechanisms to Therapeutics

“…The loss of a cells loses defining characteristics is reflected in their EVs as well. This was demonstrated by Chen et al described earlier where the ability of EVs from low passage DPCs to stimulate hair growth in ORSCs and HMCs was lost at higher passage numbers ( Chen et al, 2020b ).…”

“…Further experiments showed that miR-140-5p targeted BMP2 expression; treatment with an miR-140-5p inhibitor abolished hair growth effects. This study showed that miR-140-5p from low passage number DPC-EVs impacts HF growth and hair cycling ( Chen et al, 2020b ). EVs derived from DPCs (DPC-EVs) on hair inductive gene expression in adipose derived stem cells (ADSCs)was investigated.…”

“…bFGF present in EVs activates the FGFR downstream pathway by activating MEK and ERK1/2 and targeting bFGF downstream in DPCs ( Hu et al, 2020 ). EVs from human DPCs were shown to contain miR-140-5p that directly targeted BMP2 by downregulating its expression and subsequently increasing the transcription and translation of β-catenin, Lef-1, SHH, and GLI1 in ORSCs ( Chen et al, 2020b ). Finally, another study showed that mouse DPC-derived EVs carry hair growth-related miRNAs such as miR-195-5p, miR-214-5p, and miR-218-5p and that they increased the expression of versican, α-SMA, N-Cam, and osteopontin in APCs.…”

The Roles of Non-coding RNA in the Development and Regeneration of Hair Follicles: Current Status and Further Perspectives

“…miRNA-140-5p is enriched in low-passage DPC exosomes and targets BMP-2, which promotes ORS and matrix cell proliferation. This suggests a potential clinical application of exosomes ( Chen et al, 2020 ).…”

Editorial: Hair Follicle Stem Cell Regeneration in Aging

“…They used RNA-sequencing to identify that extracellular matrix synthesis is increased in 3D culture DP cells, and set up a transcription factor–miRNA coregulatory network in this co-culture system, which may be beneficial for future AGA studies at the molecular level. In addition to the extracellular matrix, the same group also identified that miR-140-5p in small extracellular vesicles secreted by DP cells suppresses Bmp2 expression to promote proliferation of ORS and hair matrix cells in a co-culture model ( Chen et al, 2020 ). Tissue engineering models have been established to promote hair follicle neogenesis.…”