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RNA-Seq of Epi-SC and SKPs. a A schematic diagram depicting sample preparation for RNA-Seq. Total RNA was extracted from Epi-SCs-tdTomato and SKPs-EGFP which were cultured separately or in mixture in Matrigel for 24 h. Cells were recovered from the matrix and separated by cell sorting using a flow cytometer. Total RNA was extracted from the cells and subjected to RNA-Seq analysis. b Transcriptome shows different gene transcriptional patterns of Epi-SCs and SKPs when cultured alone or in mixture (x). c, d KEGG analysis revealed active signals in Epi-SCs and SKPs when cultured in mixture, among them there was PI3K/Akt signal
Source publication
Background:
Cultured epidermal stem cells (Epi-SCs) and skin-derived precursors (SKPs) were capable of reconstituting functional hair follicles after implantation, while the signaling pathways that regulate neogenic hair follicle formation are poorly investigated. In this study, we aimed to understand the interactions between Epi-SCs and SKPs duri...
Citations
... Additionally, focal adhesion in the maintenance of hair follicle stem cell quiescence [42], and the hair follicle stem cell proliferation [43]. The PI3K-Akt signaling pathway has been extensively studied for its role in hair growth and skin development, including hair follicle regeneration [44], , hair follicle stem cell proliferation and differentiation [45], and hair-inducing dermal papilla cells [46]. Furthermore, we found some hair length-related genes, hair follicle cycle-related genes, and hair follicle growth-related genes were differently expressed between LHY and NHY. ...
Background
The hair follicle development process is regulated by sophisticated genes and signaling networks, and the hair grows from the hair follicle. The Tianzhu white yak population exhibits differences in hair length, especially on the forehead and shoulder region. However, the genetic mechanism is still unclear. Isoform sequencing (Iso-seq) technology with advantages in long reads sequencing. Hence, we combined the Iso-seq and RNA-seq methods to investigate the transcript complexity and difference between long-haired yak (LHY) and normal-haired yak (NHY).
Results
The hair length measurement result showed a significant difference between LHY and NHY on the forehead and the shoulder (P-value < 0.001). The skin samples from the forehead and the shoulder of LHY and NHY were pooled for isoform sequencing (Iso-seq). We obtained numerous long transcripts, including novel isoforms, long non-coding RNA, alternative splicing events, and alternative polyadenylation events. Combined with RNA-seq data, we performed differential isoforms (DEIs) analysis between LHY and NHY. We found that some hair follicle and skin development-related DEIs, like BMP4, KRT2, IGF2R, and COL1A2 in the forehead skin; BMP1, KRT1, FGF5, COL2A1, and IGFBP5 in the shoulder skin. Enrichment analysis revealed that DEIs in both two comparable groups significantly participated in skin and hair follicle development-related pathways, like ECM-receptor interaction, focal adhesion, and PI3K-Akt signaling pathways. The results indicated that the hair follicle development of Tianzhu white yak may influence the hair length difference. Besides, the protein-protein interaction (PPI) network of DEIs showed COL2A1 and COL3A1 exhibited a high degree of centrality, and these two genes were suggested as potential candidates for the hair length growth of Tianzhu white yak.
Conclusions
The results provided a comprehensive analysis of the transcriptome complexity and identified differential transcripts that enhance our understanding of the molecular mechanisms underlying the variation in hair length growth in Tianzhu white yak.
... The screened differential genes were input as the KEGG database for the KEGG enrichment pathway analysis, and some upregulated (Fig. 4B) and downregulated pathways (Fig. 4C) were obtained. we focused on the PI3K-Akt signaling pathway because many studies have indicated that this pathway is involved in dermal papilla-induced HFSC regeneration (Kang et al. 2020;Zhang et al. 2020;Cai et al. 2018;Wang et al. 2017;Chen et al. 2020). Thus, activation of the PI3K-Akt signaling could be a major Table 2). ...
... Indeed, inhibition of the PI3K/Akt signaling pathway can block the transformation from the S-phase to the G-phase of hair follicles (Wang et al. 2017). Prohibition of PI3K or Akt by specific inhibitors can significantly inhibit hair-follicle regeneration mediated by epidermal stem cells and skin-derived precursor cells (Chen et al. 2020). Zhang et al. (2020) showed that the vascular endothelial growth factor (VEGF) can protect CD200 + -and CD34 + -HFSCs from androgen-induced apoptosis via the PI3K/Akt signaling pathway. ...
A previous study indicated that patients with androgenic alopecia (AGA) have significantly reduced levels of LncRNA RP11-818O24.3. This study investigates whether LncRNA RP11-818O24.3 promotes hair-follicle recovery and its possible mechanism. Hair alteration and cutaneous histopathological changes induced by testosterone propionate were observed by H&E and bromodeoxyuridinc (BrdU) stain to evaluate the therapeutic effect of LncRNA RP11-818O24.3 in C57BL/6 J mice. The cellular viability was analyzed in LncRNA RP11-818O24.3-transfected human hair-follicle stem cells (HFSCs) in vitro. The signaling pathways and pro-proliferative factors were investigated by transcriptomic gene sequencing and qRT-PCR. LncRNA RP11-818O24.3 transfection successfully recovered hair growth and hair-follicle cells in AGA mice. In a series of HFSC studies in vitro, LncRNA RP11-818O24.3 transfection greatly promoted cellular proliferation and decreased cellular apoptosis. Transcriptome gene sequencing suggested that the phosphatidylinositol 3-kinase (PI3K)-Akt pathway was upregulated by LncRNA RP11-818O24.3. The qRT-PCR results showed that fibroblast growth factor (FGF)-2 was 14-times upregulated after LncRNA RP11-818O24.3 transfection. Hair-follicle recovery activity of LncRNA RP11-818O24.3 may involve the upregulation of FGF2 and PI3K-Akt to promote follicle stem cell survival. These data not only provide a theoretical basis for AGA development but also reveal a novel therapeutic method for AGA patients.
... Studies have shown that the PI3K-Akt signaling pathway plays a key role in hair follicle regeneration. PI3K inhibition significantly inhibits the hair follicle regeneration mediated by epidermal stem cells and skin-derived precursors [23,24]. ...
Background
Androgenetic alopecia (AGA) is the most common form of hair loss. Studies have suggested a potential link to metabolic disorders, but with conflicting results. To elucidate the lipidomics profile and sex-specific variations in AGA, while exploring correlation between AGA and metabolic syndrome (MetS).
Methods
The AGA patients (n = 83) and healthy controls (n = 84) were collected in the study. The lipid profiles were analyzed using ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Serum levels of important factors associated with AGA, namely dihydrotestosterone (DHT), prostaglandin D2 (PGD2) and transforming growth factor-β1 (TGF-β1) were quantified using ELISA.
Results
Compared with controls, AGA patients had a higher probability of MetS (26.51% vs 11.9%, P < 0.05). Fifty-one differentially expressed lipids were identified in AGA. The kind of triglyceride (TG) were significantly increased, while phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylinositol (PI), and phosphatidylserine (PS) exhibited remarkable decrease. PC (16:2/21:6), PC (34:4p), PE (41:7), PE (44:12), PG (40:9), PI (32:2) and TG (15:0/18:1/18:1) were identified as potential biomarkers of AGA with the highest specificity. The levels of DHT, PGD2 and TGF-β1 were significantly elevated in AGA. All seven lipids showed significant correlations with DHT, PC (34:4p) and TG (15:0/18:1/18:1) were significantly associated with PGD2, TGF-β1 displayed exclusively correlation with TG (15:0/18:1/18:1) (all P < 0.05). Furthermore, these lipids were also significantly linked to systolic blood pressure and BMI, while some of them also showed significant associations with total cholesterol and HDL-C. In subgroups, forty-two differentially expressed lipids were identified in male AGA vs male control and eighty-one in female AGA vs female control. PC (16:2/21:6) was the only specific lipids common to both sexes.
Conclusions
Aberrant lipid metabolism was observed in AGA, with distinct lipidomic profiles between male and female AGA. The potential biomarkers were closely related to DHT, PGD2, TGF-β1 and MetS-related indicators. It provides the foundation for revealing the mechanisms of AGA.
... The only metabolite enriched in this pathway was L-arginine, which has been shown to have antiaging effects on skin [47]. In our previous proteomic study, the PI3K/Akt signalling pathway was enriched, and this pathway was shown to be a potential regulatory pathway involved in hair follicle regeneration [47][48][49]. At least ten pathways related to amino acids and protein metabolism were enriched in the fine versus coarse cashmere comparison. ...
... It has been shown that the PI3K/Akt signaling pathway is essential for the morphogenesis of hair follicles. The PI3K-Akt pathway inhibitor treatment in hair follicle reconstruction experiments results in failure of follicle regeneration 30 . Additionally, our previous transcriptome sequencing results showed that the PI3K-Akt pathway was in a repressed state in HL mouse skin 4 . ...
Hair loss disorders such as androgenetic alopecia have caused serious disturbances to normal human life. Animal models play an important role in exploring pathogenesis of disease and evaluating new therapies. NIH hairless mice are a spontaneous hairless mouse discovered and bred in our laboratory. In this study, we resequenced the genomes of NIH normal mice and NIH hairless mice and obtained 3,575,560 high-quality, plausible SNP loci and 995,475 InDels. The Euclidean distance algorithm was used to assess the association of SNP loci with the hairless phenotype, at a threshold of 0.62. Two regions of chromosome 18 having the highest association with the phenotype contained 345 genes with a total length of 13.98 Mb. The same algorithm was used to assess the association of InDels with the hairless phenotype at a threshold of 0.54 and revealed a region of 25.45 Mb in length, containing 518 genes. The mutation candidate gene Lama3 (NM_010680.2: c.652C>T; NP_034810.1: p. Arg217Cys) was selected based on the results of functional gene analysis and mutation prediction screening. Lama3 (R217C) mutant mice were further constructed using CRISPR/Cas9 technology, and the relationship between Lama3 point mutations and the hairless phenotype were clarified by phenotypic observation. The results showed that male Lama3 point mutation mice started to lose hair on the 80th day after birth, and the hair loss area gradually expanded over time. H&E staining of skin sections showed that the point mutation mice had increased sebaceous glands in the dermis and missing hair follicle structure (i.e., typical symptoms of androgenetic alopecia). This study is a good extension of the current body of knowledge about the function of Lama3, and the constructed Lama3 (R217C) mutant mice may be a good animal model for studying androgenetic alopecia.
... Of the numerous signals, the simultaneous upregulation of the canonical Wnt/b-catenin signaling pathway is essential for maintaining the hair-inductive activity of DP cells [11]. Several studies have also reported that the proliferation and migration of DP cells which affect the regeneration and growth of HFs can be upregulated by the activation of ERK and AKT, and stimulation of the phosphoinositide 3-kinase (PI3K)/AKT pathway [12,13]. Therefore, numerous studies have frequently utilized DP cells for investigating the mechanism underlying the hair cycle and for the development of drugs for inducing hair growth [14]. ...
In cancer treatment, multi-target approach has paid attention to a reasonable strategy for the potential agents. We investigated whether (E)-2-methoxy-4-(3-(4-methoxyphenyl) prop-1-en-1-yl) phenol (MMPP) could exert an anticancer effect by dual-regulating VEGFR2 and PPARγ. MMPP showed modulating effects in TNBC type (MDA-MB-231 and MDA-MB-468) and luminal A type (MCF7) breast cancer cell lines. MMPP enhanced PPARγ transcriptional activity and inhibited VEGFR2 phosphorylation. MMPP-induced signaling by VEGFR2 and PPARγ ultimately triggered the downregulation of AKT activity. MMPP exhibited anticancer effects, as evidenced by growth inhibition, inducement of apoptosis, and suppression of migration and invasion. At the molecular level, MMPP activated pro-apoptotic proteins (caspase3, caspase8, caspase9, and bax), while inhibiting the anti-apoptotic proteins (bcl2). Additionally, MMPP inhibited the mRNA expressions of EMT-promoting transcription factors. Therefore, our findings showed molecular mechanisms of MMPP by regulating VEGFR2 and PPARγ, and suggested that MMPP has potential to treat breast cancer.
... Indicating the importance of 3D cultures, appropriate EMI's from co-culture and low passage number [74]. Furthermore, de novo HFs were generated, when a mouse epidermal and dermal cell matrix was transplanted into an in vivo mouse wound model [88]. Indicating that organoids require additional signals from the native tissue environment and more research is required relating to the signalling pathways from other cell populations. ...
... The PI3K/AKT signalling pathway is another important regulator in the cross talk between epithelial and mesenchymal cells for HF growth. Similarly, the PI3K/AKT signalling pathway is required for telogen to anagen phase progression and stimulates proliferation in HF stem cells [88]. In brief a relevant ligand activates PI3K (phosphoinositol 3-kinase) and tiggers the phosphorylation and activation of AKT, AKT affects glucose metabolism and promotes protein synthesis, proliferation and differentiation of HFSCs [103]. ...
Androgenic alopecia is a hereditary condition of pattern hair loss in genetically susceptible individuals. The condition has a significant impact on an individual's quality of life, with decreased self-esteem, body image issues and depression being the main effects. Various conventional treatment options, such as minoxidil, finasteride and herbal supplements, aim to slow down hair loss and promote hair growth. However, due to the chronic nature of the condition the financial cost of treatment for androgenic alopecia is very high and conventional treatment options are not universally effective and come with a host of side effects. Therefore, to address the limitations of current treatment options a novel regenerative treatment option is required. One promising approach is organoids, organoids are 3D cell aggregates with similar structures and functions to a target organ. Hair follicle organoids can be developed in vitro. However, the main challenges are to maintain the cell populations within the organoid in a proliferative and inductive state, as well as to promote the maturation of organoids. Photobiomodulation is a form of light therapy that stimulates endogenous chromophores. PBM has been shown to improve cell viability, proliferation, migration, differentiation and gene expression in dermal papilla cells and hair follicle stem cells. Therefore, photobiomodulation is a potential adjunct to hair follicle organoid culture to improve the proliferation and inductive capacity of cells.
... Previous studies used transgenic mouse that marked the epithelial nuclei of skin by expressing a fusion protein of histone H2B with green fluorescent protein driven by the keratin 14 promoter (K14H2BGFP) [35,36], TPEF imaging detected hair follicle by marking the epithelial nuclei. To investigate neogenic hair follicle formation, epidermal stem cells (Epi-SCs) derived from neonatal C57BL/6 mice were labelled with tdTomato, and skin-derived precursors (SKPs) were isolated from neonatal C57BL/6/GFP mice, and reconstituting function of hair follicles from Epi-SCs and SKPs was tracked using TPEF imaging [37]. TRITC-conjugated dextran/FITC-conjugated dextran was injected into the tail vein to evaluate blood vessel leakage using TPEF imaging [38]. ...
The identification of crucial targets for hair regrowth in androgenetic alopecia (AGA) involves determining important characteristics and different stages during the process of hair follicle regeneration. Traditional methods for assessing key features and different stages of hair follicle primarily involve taking skin tissue samples and determining them through various staining or other methods. However, non-invasive assessment methods have been long sought. Therefore, in this study, endogenous fluorescence signals from skin keratin and second harmonic signals from skin collagen fibers were utilized as probes, two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) imaging techniques were employed to non-invasively assess hair shafts and collagen fibers in AGA mice in vivo. The TPEF imaging technique revealed that the alternation of new and old hair shafts and the different stages of the growth period in AGA mice were delayed. In addition, SHG imaging found testosterone reduced hair follicle area and miniaturized hair follicles. The non-invasive TPEF and SHG imaging techniques provided important methodologies for determining significant characteristics and different stages of the growth cycle in AGA mice, which will facilitate future non-invasive assessments on human scalps in vivo and reduce the use of animal testing.
... It was postulated that these pathways play an essential role in the onset of anagen, keratinocyte growth, and differentiation as well as hair follicle regeneration by either positive or negative Frontiers in Genetics frontiersin.org 11 feedback mechanisms (Bernard, 2003;Calautti et al., 2005;Harel et al., 2015;Chen et al., 2020). In particular, the differentiation of keratinocytes, their initiation of rapid mitosis and/or keratinization were suspected to be important to determine the curly hair fiber shape (Thibaut et al., 2005;Thibaut et al., 2010;Sriwiriyanont et al., 2011). ...
Hair types have been under strong targeted selection in domestic animals for their impact on skin protection, thermoregulation and exterior morphology, and subsequent economic importance. In pigs, a very special hair phenotype was observed in Mangalitza, who expresses a thick coat of curly bristles and downy hair. Two breed-specific missense variants in TRPM2 and CYP4F3 were suggested to be associated with the Mangalitza pig’s hair shape due to their role in hair follicle morphogenesis reported for human and mice. However, the mechanism behind this expression of a curly hair type is still unclear and needs to be explored. In our study, hair shafts were measured and investigated for the curvature of the hair in Mangalitza and crossbreeds in comparison to straight-coated pigs. For molecular studies, hair roots underwent RNA sequencing for a differential gene expression analysis using DESeq2. The output matrix of normalized counts was then used to construct weighted gene co-expression networks. The resulting hair root gene expression profiles highlighted 454 genes to be significantly differentially expressed for initiation of curly hair phenotype in newborn Mangalitza piglets versus post-initiation in later development. Furthermore, 2,554 genes showed a significant differential gene expression in curly hair in comparison to straight hair. Neither TRPM2 nor CYP4F3 were identified as differentially expressed. Incidence of the genes in weighted co-expression networks associated with TRPM2 and CYP4F3, and prominent interactions of subsequent proteins with lipids and calcium-related pathways suggested calcium signaling and/or lipid metabolism as essential players in the induction of the curly hair as well as an ionic calcium-dependency to be a prominent factor for the maintenance of this phenotype. Subsequently, our study highlights the complex interrelations and dependencies of mutant genes TRPM2 and CYP4F3 and associated gene expression patterns, allowing the initiation of curly hair type during the development of a piglet as well as the maintenance in adult individuals.
... Hydrangea has been reported to promote hair regrowth and repair in mouse models of hair loss through activation of TGF-β signaling and mitochondrial signaling pathways 46 . The PI3K-AKT signaling pathway is critical to human hair follicle regeneration 47 . In addition, PI3K-AKT, MAPK, Ras and Rap1 signaling pathways play a certain role in the growth of in vitro cultured Inner Mongolian cashmere goat hair follicle stem cells 48 . ...
The Liaoning cashmere goat has been confirmed as a valuable genetic resource breed that is prohibited from genetic outflow in China, and it achieves the highest single fleece production. Hair follicle development in the cashmere goat is regulated by melatonin and long non-coding RNAs (lnRNAs). However, the role played by lncRNAs in mediating melatonin-promoted cash-mere growth remains unclear. A novel lncRNA-lncRNA018392 with significant overexpression, which played a certain role in the melatonin-promoted proliferation of cashmere skin fibroblasts, was screened in previous research. The flow cytometry and CCK-8 results confirmed that the knockdown of lncRNA018392 reversed the effect of melatonin on cell proliferation, and the prolif-eration of cashmere skin fibroblasts was inhibited after the cells were interfered with the gene CSF1R near lncRNA018392. The dual-luciferase reporter assay further demonstrated that lncRNA018392 can positively regulate the promoter of CSF1R. Moreover, as indicated by the results of RNA-binding protein immunoprecipitation (RIP) and chromatin immunoprecipitation sequencing (ChIP-Seq), lncRNA018392 formed a complex with transcription factor SPI1, and CSF1R served as a downstream target gene regulated by SPI1. As revealed by the results of this study, melatonin-mediated novel lncRNA018392 accelerated the cell cycle, facilitated cell proliferation, and inhibited apoptosis by recruiting SPI1 to up-regulate the expression of nearby gene CSF1R. This study lays a theoretical basis for clarifying the molecular mechanism of cashmere growth and molecular breeding of cashmere goats.
