Hair Follicle Stem Cells Research Articles

Melatonin-Mediated Circadian Rhythm Signaling Exhibits Bidirectional Regulatory Effects on the State of Hair Follicle Stem Cells

The development and regulation of hair are widely influenced by biological rhythm signals. Melatonin plays a crucial role as a messenger in transmitting biological rhythm signals, and its impact on hair development has been well documented. During the process of hair follicle reconstruction, hair follicle stem cells (HFSCs) are the most important cell type, but the regulatory effect of melatonin on the state of HFSCs is still not fully understood. Therefore, it is necessary to conduct a more comprehensive characterization of the effects of melatonin on the state of hair follicle stem cells. The research results indicate that HFSCs express retinoic acid receptor-related orphan receptor alpha (Rorα), and melatonin inhibits the expression level of RORA. Experimental results from CUT&Tag, CUT&RUN, and dual luciferase reporter assays demonstrate that Foxc1 is a downstream target gene of RORA, with RORA regulating Foxc1 expression by binding to the promoter region of Foxc1. The CCK-8 assay results show that low doses of melatonin upregulate the survival rate of hair follicle stem cells, while high doses have the opposite effect. The knockdown of Foxc1 reverses the inhibitory effect of high-dose melatonin on the survival rate of hair follicle stem cells. Based on these findings, we believe that melatonin-mediated circadian signals exert a bidirectional regulatory effect on the state of HFSCs.

Dahuang-Gancao decoction ameliorates testosterone-induced androgenetic alopecia in mice.

Dahuang-Gancao decoction (DGD) is a traditional Chinese medicinal formula that is recorded in the Synopsis of the Golden Chamber, and is widely used to treat damp-heat in the body. Since the pathological factors of androgenetic alopecia (AGA) also reflect damp-heat blockage, DGD has great potential for the treatment of AGA and has been used effectively in clinical practice. The aim of the study was to investigate whether external application of DGD could promote the activation and proliferation of hair follicle stem cells (HFSCs) and improve AGA through the Wnt/β-catenin pathway. The main chemical components of DGD-contained serum were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and database search. Cell Counting Kit-8 (CCK8) was used to investigate the appropriate concentration. Hair regeneration was assessed by hair growth score and histopathological staining. The proliferation of HFSCs and the activation of Wnt/β-catenin pathway were detected by Western blot, immunofluorescence staining, real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and enzyme-linked immunosorbent assay (ELISA). The AGA mouse model was induced by external application of testosterone (T). Immunofluorescence staining was performed to localize HFSCs by CK15, followed by staining with Ki67, β-catenin, and Cyclin D1, respectively. The results illustrated that the 10% DGD group and the 10% DGD+HLY78 group could significantly promote the expression of Wnt10b and β-catenin and the proliferation of HFSCs in vitro, while the 10% DGD+IWR-1 group could reverse the promotion effect of DGD. Animal experiments showed that compared with the model group (T group), DGD promoted hair follicles to enter the anagen phase, as evidenced by an increase in hair growth score, an increase in the number of hair follicles in hematoxylin and eosin (HE) staining, and a significant increase in the ratio of the number of anagen follicles to the total number of hair follicles (AF/AF+TF). In addition, DGD upregulated the expression of Wnt/β-catenin signaling pathway proteins in the skin tissues of AGA mice. It also promoted the proliferation of HFSCs and the expression of β-catenin and Cyclin D1 cytokines in the region of HFSCs. Both oral and external application of DGD can promote the proliferation of HFSCs by activating the Wnt/β-catenin signalling pathway. External application of DGD can promote the hair follicles to enter the anagen phase, which can ameliorate the symptoms of alopecia in AGA mice. Therefore, compared to oral DGD, external application of DGD is an effective and safer way of administration for the treatment of AGA.

Sphingolipid metabolism orchestrates establishment of the hair follicle stem cell compartment.

Sphingolipids serve as building blocks of membranes to ensure subcellular compartmentalization and facilitate intercellular communication. How cell type-specific lipid compositions are achieved and what is their functional significance in tissue morphogenesis and maintenance has remained unclear. Here, we identify a stem cell-specific role for ceramide synthase 4 (CerS4) in orchestrating fate decisions in skin epidermis. Deletion of CerS4 prevents the proper development of the adult hair follicle bulge stem cell (HFSC) compartment due to altered differentiation trajectories. Mechanistically, HFSC differentiation defects arise from an imbalance of key ceramides and their derivate sphingolipids, resulting in hyperactivation of noncanonical Wnt signaling. This impaired HFSC compartment establishment leads to disruption of hair follicle architecture and skin barrier function, ultimately triggering a T helper cell 2-dominated immune infiltration resembling human atopic dermatitis. This work uncovers a fundamental role for a cell state-specific sphingolipid profile in stem cell homeostasis and in maintaining an intact skin barrier.

Pathogenesis and regenerative therapy in vitiligo and alopecia areata: focus on hair follicle.

Vitiligo is an autoimmune disease characterized by the loss of functional melanocytes in the hair follicles and epidermis, leading to white patches on the skin and mucous membranes. Alopecia areata (AA) is a common immune-mediated condition in which autoimmune attack on hair follicles cause non-scarring hair loss. Both diseases significantly impact patients's physical and mental health. Hair follicles, dynamic mini-organs, house diverse stem cell populations that form hair structures. Melanocyte stem cell (McSCs) and hair follicle stem cells (HFSC) located in the hair follicle bulge contribute to follicular structures during each anagen phase of the hair cycle, synchronizing periodic activities to impact color to the hair. Hair follicle dysfunction may contribute to hair loss and could potentially interfere with repigmentation efforts in vitiligo lesions. This article reviews the role of hair follicles in the pathogenesis, clinical manifestations, and therapeutic options for vitiligo and AA, aiming to deepen clinicians' understanding of follicular involvement in these diseases and explore potential treatment avenues.

BMP4 regulates differentiation of nestin-positive stem cells into melanocytes

Hair follicle (HF) development and pigmentation are complex processes governed by various signaling pathways, such as TGF-β and FGF signaling pathways. Nestin + (neural crest like) stem cells are also expressed in HF stem cells, particularly in the bulge and dermal papilla region. However, the specific role and differentiation potential of these Nestin-positive cells within the HF remain unclear, especially regarding their contribution to melanocyte formation and hair pigmentation. Bone morphogenetic protein 4 (BMP4), members of the TGFβ family, has been implicated in regulating HF growth, coloration, and related cellular behaviors. Its role in directing Nestin-positive cells toward a melanocytic lineage has yet to be fully explored. In this study, mouse HF organoids were constructed and shown to be an ideal model for studying HF growth and development in vitro. Using this model as a basis, we demonstrated that BMP4 controls HF coloration as well as its length, number, and even size. Furthermore, Nestin-positive cells in the HF-especially those in the bulge region-differentiate into melanocytes, which produce the pigments that give HF its color under BMP4 stimulation. The resulting increase in pigmentation within the mouse HF organoids underscores that BMP4 has a major regulatory role in the formation of melanocytes from Nestin-positive stem cells. This research provides insights into the cellular mechanisms underlying hair pigmentation and suggests potential therapeutic applications for pigmentation disorders.

Retinoic acid drives hair follicle stem cell activation via Wnt/β-catenin signalling in androgenetic alopecia.

Depletion or permanent quiescence of the hair follicle stem cell (HFSC) pool underlies pathogenesis in androgenetic alopecia (AGA). Reactivation of quiescent HFSCs is considered an efficient treatment strategy for hair loss. The retinoic acid (RA) is critical to ensure stem cell homeostasis and function. However, little is known about whether RA regulates HFSC homeostasis. We aimed to investigate the impact of RA on HFSC homeostasis and the underlying mechanisms, in order to provide new potential targets for medical therapies of AGA. Microdissected hair follicles from the occipital and frontal scalp in AGA were obtained for RNA sequencing analysis and test. The C57BL/6 mice model in telogen was established to investigate the effect of exogenous RA. Miniaturized hair follicles from frontal scalp were incubated with or without RA in hair follicle organ culture to test the effects on hair shaft elongation, hair cycling and HFSC activities. A strategy to characterize the effect of RA on HFSC in primary culture was developed to identify novel mechanisms that control HFSC activation. A clinical study was performed to test the efficacy of RA treatment in AGA patients. RA signalling was inhibited in the course of AGA pathogenesis along with HFSC dysfunction. Hair regeneration was retarded in AGA miniaturized hair follicles with RA deficiency, but they tended to recover after treatment with RA. In addition, RA treatment during the telogen phase facilitated HFSC anagen entry and accelerated hair growth. Mechanistically, RA promoted hair growth by stimulating stem cells via Wnt/β-catenin signalling and accelerating the transition from a dormant to an activated state. Furthermore, a clinical study suggested that RA has obvious advantages in the early intervention of AGA by reactivating HFSCs. Our study provides insights into the reactivation of HFSCs in AGA and provides potential targets for medical therapies.

FGF20 Secreted From Dermal Papilla Cells Regulate the Proliferation and Differentiation of Hair Follicle Stem Cells in Fine-Wool Sheep.

Wool traits determine the market value of fine-wool sheep, and wool fibre-breaking elongation (fibres can be stretched or elongated before they break) is one of the important wool traits. The interaction between hair follicle stem cells (HFSCs) and dermal papilla cells (DPCs) determines hair follicle development in fine wool sheep, thereby directly influencing wool traits. A genome-wide association study based on pre-sequencing data identified FGF20, which was significantly associated with wool fibre-breaking elongation. The study reveals that the regulatory mechanism of FGF20 secreted from DPCs affects the proliferation and differentiation of HFSCs through a co-culture system, to provide a new perspective for fine-wool sheep breeding. After knocking down FGF20 expression in DPCs, the results showed that the expression of fibroblast growth factor receptor 2 (FGFR2) and fibroblast growth factor receptor 3 (FGFR3) in DPCs and HFSCs was significantly decreased (p < 0.05), the number of EdU-positive cells and cell viability was significantly decreased (p < 0.01), and the apoptosis rate was significantly increased (p < 0.05). Meanwhile, the differentiation markers of SOX9, NOTCH1 and β-Catenin in HFSCs were also significantly reduced (p < 0.05). These findings indicate that FGF20-knockdown in DPCs of fine-wool sheep inhibits the proliferation and differentiation of HFSCs in the co-culture system, providing a theoretical basis for elucidating the regulatory mechanism of hair follicle self-renewal and differentiation of fine-wool sheep and providing a co-culture system for regenerative medicine.

Isolation, Expansion, and Characterization of Rat Hair Follicle Stem Cells and Their Secretome: Insights into Wound Healing Potential.

Background: Stem cells are capable of self-renewal and differentiation into various specialized cells, making them a potential therapeutic option in regenerative medicine. This study establishes a comprehensive methodology for isolating, culturing, and characterizing rat hair follicle stem cells. Methods and Results: Hair follicles were harvested from Sprague-Dawley rats and subjected to two different isolation techniques. Immunohistochemical analysis and real-time PCR confirm the expression of specific surface markers and genes, validating the cells' identity. Growth kinetics, colony formation units (CFU), and tri-differentiation capacity were also assessed. Additionally, the cells' secretome was analyzed, regarding its content in biofactors with wound healing properties. Conclusions: These findings highlight the potential of these cells as a valuable cell source for skin regeneration applications. They contribute to advancing our understanding of stem cell applications in regenerative medicine and hold promise for therapeutic interventions in various clinical contexts, aligning with broader research on the diverse capabilities of hair follicle stem cells.

In vitro hair growth-promoting effects of araliadiol via the p38/PPAR-γ signaling pathway in human hair follicle stem cells and dermal papilla cells.

Scalp hair plays a crucial role in social communication by expressing personal appearance and self-identity. Consequently, hair loss often leads to a perception of unattractiveness, negatively impacting an individual's life and mental health. Currently, the use of Food and Drug Administration (FDA)-approved drugs for hair loss is associated with several side effects, highlighting the need for identifying new drug candidates, such as plant-derived phytochemicals, to overcome these issues. This study investigated the hair growth-promoting effects of araliadiol, a polyacetylene compound found in plants such as Centella asiatica. We employed an in vitro model comprising human hair follicle stem cells (HHFSCs) and human dermal papilla cells (HDPCs) to evaluate the hair growth-promoting effects of araliadiol. The proliferation-stimulating effects of araliadiol were assessed using water-soluble tetrazolium salt assay, adenosine triphosphate content assay, and crystal violet staining assay. In addition, we performed luciferase reporter assay, polymerase chain reaction analysis, cell fractionation, Western blot analysis, and enzyme-linked immunosorbent assay (ELISA) to elucidate the mechanism underlying the hair growth-inductive effects of araliadiol. Araliadiol exhibited both proliferation- and hair growth-promoting effects in HHFSCs and HDPCs. Specifically, it increased the protein expression of cyclin B1 and Ki67. In HHFSCs, it elevated the expression of hair growth-promoting factors, including CD34, vascular endothelial growth factor (VEGF), and angiopoietin-like 4. Similarly, araliadiol increased the expression of hair growth-inductive proteins such as fibroblast growth factor 7, VEGF, noggin, and insulin-like growth factor 1 in HDPCs. Subsequent Western blot analysis and ELISA using inhibitors such as GW9662 and SB202190 confirmed that these hair growth-promoting effects were dependent on the p38/PPAR-γ signaling in both HHFSCs and HDPCs. Araliadiol promotes hair growth through the p38/PPAR-γ signaling pathway in human hair follicle cells. Therefore, araliadiol can be considered a novel drug candidate for the treatment of alopecia.

Retinoic Acid-Related Orphan Receptor Alpha May Regulate the State of Hair Follicle Stem Cells by Upregulating the Expression of BNIP3.

The hair, an exclusive keratinized dermal appendage in mammals, stands as a quintessential outcome of adaptive evolution, conferring resilience against adverse environmental conditions. The ontogenesis of the coat displays a pronounced rhythmic pattern, with hair follicle stem cells (HFSCs) emerging as pivotal facilitators of hair follicle reconstitution. The retinoic acid-related orphan receptor alpha, a nuclear receptor with extensive involvement in the regulation of cellular physiological states, exerts its functions predominantly through the modulation of downstream target gene transcription. The Bnip3 gene exhibits a robust correlation with cellular apoptosis and autophagy, which are indispensable physiological mechanisms underlying the maintenance of HFSC homeostasis. Consequently, the expression level of Bnip3 may be intimately linked to the status of HFSCs. In this investigative endeavor, we employed rat HFSCs as a model system to validate the regulatory impact of RORA on Bnip3 gene expression. Our findings unequivocally demonstrate that Bnip3 serves as a direct downstream target of RORA. Specifically, RORA binds to the motif within the Bnip3 promoter region, thereby upregulating Bnip3 expression levels. In light of our research findings, we propose that RORA holds potential as a target for modulating the status of HFSCs.

Effects of Petasitin as Natural Extract on Proliferation and Pathological Changes of Pediatric Neuroblastoma SK-N-SH Cells

Scalp alopecia areata (SAA) is a common non-scarring hair loss condition, associated with factors such as autoimmune responses, genetics, emotional stress, and endocrine imbalances. Current treatments for SAA included minoxidil, topical steroid creams, biologics, and plant extracts. Tea tree oil (TTO), a natural plant extract, is known for its antibacterial, anti-inflammatory, and acaricidal properties, and it also provides nourishment for hair. In this research, a natural extract of TTO, was prepared to analyze its antibacterial properties. The hair follicle stem cells (HFSCs) of patients with primary SAA were analyzed to understand the influences of TTO on migration of HFSCs. TTO was extracted from fresh tea tree leaves using steam distillation. Quantitative analysis of gas chromatography-mass spectrometry (GC/MS) qualitative analysis and its total ion chromatogram using area normalization method were conducted. Meanwhile, its antibacterial activity was tested against five common pathogens (Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), Staphylococcus albus (S. albus), Pseudomonas aeruginosa (P. aeruginosa), and Candida albicans (C. albicans)) by measuring the diameter of inhibition zones (DIZ), minimal inhibitory concentration (MIC), and minimum bactericidal concentration (MBC). HFSCs were isolated from patients with SAA and cultured in vitro, with cell identification performed through cytokeratin 15 (K15) immunofluorescent staining. The HFSCs were then exposed to varying concentrations (0.0, 0.5, 2.0, 5.0, 10.0, and 25.0 mmol/L) of TTO for culture, and cell proliferation activity (CPA) was assessed using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay, while migration of HFSCs was evaluated using the Transwell chamber assay. Results demonstrated that the extracted TTO had a content of 0.69 g and an extraction rate of 2.32%. 36 components were identified, constituting 98.67% of the total, with 4-terpineol reaching a high concentration of 48.35%. It exhibited a DIZ of less than 25 mm against all tested pathogens, with MIC values lower than 29 mg/mL and MBC values below 38 mg/mL. Patients with SAA displayed yellow and black dots, broken hair, malnourished and exclamation mark hairs, with few flag hairs observed in skin microscope examination. Isolated and cultured HFSCs expressed K15 primarily in the cytoplasm and exhibited favorable growth dynamics. Treatment with various concentrations of TTO greatly increased CPA and migrated cell numbers in HFSCs, with the optimal effect observed at 5.0 mmol/L concentration of TTO. In conclusion, the plant extract TTO possessed significant antibacterial activity and can promote proliferation and migration in vitro of HFSCs, suggesting its potential therapeutic application for SAA.

An anti-sense lncRNA of the A-FABP gene regulates the proliferation of hair follicle stem cells via the chi-miR-335-5p/DKK1/β-catenin axis.

Hair follicle development relies on both the epithelial-mesenchymal interaction (EMI) and the proliferation of hair follicle stem cells (HFSCs). This intricate process involves numerous regulatory molecules. Increasing evidence suggests that long non-coding RNAs (lncRNAs) play a crucial role in hair follicle development. However, the functions and molecular mechanisms of many lncRNAs in hair follicle development of cashmere goats remain unclear. Based on our previous lncRNA sequencing results in cashmere goats, an unannotated lncRNA differentially expressed at various stages of hair follicle development, named FABP_AS, was detected. Consequently, we aimed at exploring the function and molecular mechanisms of FABP_AS. We constructed a CRISPR/Cas9 knockout system to specifically knock down FABP_AS, providing a reference model for target lncRNA knockout in animal primary cells. Functional experiment results demonstrated that FABP_AS significantly inhibited HFSCs proliferation. Mechanism experiment results revealed that FABP_AS competitively bond to chi-miR-335-5p, promoted DKK1 gene expression, and reduced Wnt/β-catenin signaling pathway activity. In summary, our findings indicated that FABP_AS acted as a miRNA sponge, sequestering chi-miR-335-5p away from the DKK1 gene, thereby suppressing HFSCs proliferation, which would lay the groundwork for a better understanding of the molecular mechanisms of hair follicle development and provide therapeutic targets for hair loss.

Intermittent fasting triggers interorgan communication to suppress hair follicle regeneration.

Intermittent fasting has gained global popularity for its potential health benefits, although its impact on somatic stem cells and tissue biology remains elusive. Here, we report that commonly used intermittent fasting regimens inhibit hair follicle regeneration by selectively inducing apoptosis in activated hair follicle stem cells (HFSCs). This effect is independent of calorie reduction, circadian rhythm alterations, or the mTORC1 cellular nutrient-sensing mechanism. Instead, fasting activates crosstalk between adrenal glands and dermal adipocytes in the skin, triggering the rapid release of free fatty acids into the niche, which in turn disrupts the normal metabolism of HFSCs and elevates their cellular reactive oxygen species levels, causing oxidative damage and apoptosis. A randomized clinical trial (NCT05800730) indicates that intermittent fasting inhibits human hair growth. Our study uncovers an inhibitory effect of intermittent fasting on tissue regeneration and identifies interorgan communication that eliminates activated HFSCs and halts tissue regeneration during periods of unstable nutrient supply.

Restoration of follicular β-catenin signaling by mesenchymal stem cells promotes hair growth in mice with androgenetic alopecia

BackgroundThe use of mesenchymal stem cells (MSCs) is recognized as a promising strategy for the treatment of androgenetic alopecia (AGA). However, the underlying mechanism remains to be explored. Here, we evaluated the therapeutic effects and potential mechanisms of the use of human umbilical cord mesenchymal stem cells (hUCMSCs) in dihydrotestosterone (DHT)-induced AGA models in vivo and in vitro.MethodsIntradermal transplantation of hUCMSCs was performed in AGA model mice and therapeutic effects were evaluated using histological and immunofluorescence staining. Transwell assays were used for co-culture of hUCMSCs and dermal papilla cells (DPCs), and communication was assessed using RT-qPCR, immunofluorescence, and apoptosis analysis. Interactions between DPCs and hair follicle stem cells (HFSCs) were investigated using RT-qPCR, EdU assays, and cell cycle analysis. ResultsTreatment of AGA mice with hUCMSCs promoted hair growth, HFs density, skin thickness, and anagen phase activation, while inhibiting DPCs apoptosis, and promoting HFSCs proliferation. In vitro, hUCMSCs activated Wnt/β-catenin signaling in DPCs via Wntless (Wls), while stimulating growth factor secretion and HFSCs proliferation. Blocking β-catenin degradation with MSAB increased DPCs apoptosis, reduced growth factor secretion, and retarded HFSCs proliferation.ConclusionhUCMSCs promoted hair regeneration in AGA model mice. This was found to be dependent on reducing DPCs apoptosis, thereby relieving the inhibitory effects of DPCs on the growth of HFSCs. The activation of the Wnt/β-catenin signaling pathway was shown to play a crucial role in the promotion of hair growth by hUCMSCs in AGA mice.

In Vitro differentiation of hair-follicle bulge stem cells into synaptophysin-expressing neurons: a potential new approach for neuro-regeneration.

Stem cells, particularly bulge hair follicle stem cells (HFSCs), have recently attracted significant interest due to their potential for tissue repair and regeneration. These cells, marked by their expression of Nestin (a neural stem cell marker), suggest the possibility of neural differentiation into neurons. This study investigated the use of retinoic acid (RA) and epidermal growth factor (EGF) to induce HFSC transformation into mature neurons, identified by synaptophysin expression. Rat whisker follicles were cultured in a medium suitable for HFSC survival and proliferation. Immunostaining techniques were used to identify HFSCs and assess their differentiation into neural cells. The addition of RA and EGF to the culture medium aimed to induce this differentiation. Findings demonstrate that HFSCs expressed Nestin, indicating their pluripotent nature. Treatment with RA and EGF resulted in synaptophysin expression, a marker of mature neurons, which was absent in the control group. However, this treatment group also displayed a decrease in the expression of other neural markers (βIII tubulin and NeuN). This study suggests that a combination of RA and EGF can accelerate HFSC differentiation into synaptophysin-positive cells in vitro. This research paves the way for further exploration of its potential application in neuro-regeneration.

Abstract B007: Unlocking antiviral immunity against endogenous retroviruses in skin squamous cell carcinomas

Abstract Known as molecular parasites that invaded our genome through ancestors, transposons (mainly retrotransposons) are interspersed genomic repeats that constitute ∼40% of the mammalian genome. Primarily residing in the heterochromatin, retrotransposons exhibit dynamic expressions to orchestrate embryonic development and shape adult functions. Although rare, a cohort of evolutionarily young retrotransposons resisted extinction, whose selfish activities underlie polymorphic variations and genetic diseases including cancer. While the extraordinary molecular and functional heterogeneity of retrotransposons has been long recognized, mechanistic dissection remains challenging. To tackle this challenge, we use murine skin as our model, given its well-characterized, abundant, and highly accessible adult stem cells, not only mediating postnatal remodeling and tissue repair, but also serving as cell of origin for squamous cell carcinomas. By analyzing a genetic model lacking a known retrotransposon suppressor, histone methyltransferase Setdb1, which we found to be essential in the adult skin, we saw hair loss and hair follicle stem cell exhaustion phenotype, accompanied by a robust and selective surge of endogenous retroviruses (ERVs). When combined with squamous cell carcinoma drivers, Setdb1 loss significantly blocked tumor progression in vivo. We detected abundant retroviral peptides originated from its full-length copies through mass spectrometry and viral-like particles through transmission electron microscopy that are immunoreactive to previously reported ERV surface envelope (env) antibody. Similar viral-like particles were broadly evident across several epithelial tissues beyond skin, implicating its conserved function across squamous cancers. Mechanistically, epithelial originated ERVs elicit tissue wide inflammatory response, and can be ameliorated by pharmacological or genetic inhibition of retroviral activity. Moreover, ERV reactivation induced replication stress functionally accounts for stem cell exhaustion, therefore serving as a specific therapeutic vulnerability in squamous cancers. As an evolutionary conundrum, viral-coding ERVs pose a threat to host fitness and yet, they’ve resisted extinction persisting in the mammalian genome. Our findings suggest ERVs wrestle with the host surveillance programs to regulate adult tissue physiology and pathology, thus providing a key target in eliciting innate immunity in squamous cell carcinomas. Citation Format: Ying Lyu, Yejing Ge. Unlocking antiviral immunity against endogenous retroviruses in skin squamous cell carcinomas [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: RNAs as Drivers, Targets, and Therapeutics in Cancer; 2024 Nov 14-17; Bellevue, Washington. Philadelphia (PA): AACR; Mol Cancer Ther 2024;23(11_Suppl):Abstract nr B007.

JAK-STAT1 as therapeutic target for EGFR deficiency-associated inflammation and scarring alopecia

The hair follicle stem cell niche is an immune-privileged microenvironment, characterized by reduced antigen presentation, thus shielding against permanent immune-mediated tissue damage. In this study, we demonstrated the protective role of hair follicle-specific epidermal growth factor receptor (EGFR) against scarring hair follicle destruction. Mechanistically, disruption of EGFR signaling generated a cell-intrinsic hypersensitivity within the JAK-STAT1 pathway, which, synergistically with interferon gamma expressing CD8 T-cell and NK-cell-mediated inflammation, compromised the stem cell niche. Hair follicle-specific genetic depletion of either JAK1/2 or STAT1 or therapeutic inhibition of JAK1/2 ameliorated the inflammation, restored skin barrier function and activated the residual stem cells to resume hair growth in mouse models of epidermal and hair follicle-specific EGFR deletion. Skin biopsies from EGFR inhibitor-treated and cicatricial alopecia patients revealed an active JAK-STAT1 signaling signature along with upregulation of antigen presentation and downregulation of key components of the EGFR pathway. Our findings offer molecular insights and highlight a mechanism-based therapeutic strategy for addressing chronic folliculitis associated with EGFR-inhibitor anti-cancer therapy and cicatricial alopecia.

Exosomes derived from Umbilical cord mesenchymal stem cell promote hair regrowth in C57BL6 mice through upregulation of the RAS/ERK signaling pathway.

Androgenetic alopecia is one of the common types of hair loss and has become a medical and social problem due to its increasingly young onset. Existing therapies, although effective, have serious side effects and therefore better treatments need to be sought. The aim of this study was to evaluate the efficacy of umbilical cord mesenchymal stem cell-derived exosomes in the treatment of androgenetic alopecia and to investigate the mechanism of exosome regulation of hair growth. First, we randomly divided 20 C57BL/6J mice into blank group, model group, positive control group and exosomal hydrogel group, and mice were treated with hair removal on the back. The mice were injected intraperitoneally with dihydrotestosterone solution except for the blank group. At the end of the experiment, new hairs were collected and the differences in length, diameter and number of hair follicles were compared among the groups; the histopathological changes of hair follicles were observed by HE staining; the expression of androgen receptor mRNA and protein in skin tissues were compared; and the skin tissues were analyzed by real-time PCR, western blotting, immunofluorescence staining and transcriptome sequencing. Finally, the results of transcriptome sequencing experiments were verified by real-time PCR, western blotting and other techniques for the corresponding genes and proteins. Compared with the blank group, mice in the model group had shorter hair length and reduced hair diameter, and pathological observation showed that the total number of hair follicles was significantly reduced and the hair follicles were miniaturized; compared with the model group, mice in the positive control and exosome groups had longer hair length, larger hair diameter and more hair follicles; the androgen receptor mRNA content and protein expression in the skin tissue of mice in the model group were significantly higher than those in the blank group, and the protein expression in the exosome gel group was lower than that in the model group. Similarly, compared with the model group, the expression of stemness-related proteins K15 and CD200 in the skin tissues of mice in the exosome group increased, and the expression of PCNA, a protein related to cell proliferation, increased. The KEGG data showed that the differential genes were mainly enriched in the RAS/ERK pathway. In this study, we demonstrated the therapeutic effect of umbilical cord MSC-derived exosomes on androgenetic alopecia and verified that exosomes regulate hair follicle stem cell stemness through the RAS/ERK pathway to promote hair proliferation and thus hair growth in mice with androgenetic alopecia, providing a potential therapeutic strategy for androgenetic alopecia.

Stem cells feel the pain

Pain is a sensation that signals the presence of inflammation or injury. In this issue of Developmental Cell, Ben-Shaanan etal.1 show that beyond its sensory function, pain can activate hair follicle stem cells (HFSCs) by controlling their niche.

Cardiomyocyte proliferation is promoted by intracellular Gas6-mediated Yap activity during neonatal heart regeneration

Abstract Purpose The adult mammalian heart has limited regenerative capacity after injury, mostly attributable to postnatal cardiomyocyte (CM) cell cycle arrest. The key to unlocking the regenerative potential of the adult hearts may lie within the developmental transitions occurring during neonatal life. Gas6, a secretory protein, stimulates the proliferation of various type cells through binding to TAM receptor family, such as endothelial progenitor cells, tumor cells, and hair follicle stem-cell. However, no study to date has studied the role of Gas6 during the proliferation of CMs. Herein, our study uncovers that Gas6 is transiently expressed during early postnatal heart and regenerative heart, and functions through intracellular pathway by regulating Yap activity to promote CM proliferation. Methods CM isolation was conducted to assess the change of Gas6 expression in both postnatal development and regenerated stages using the CM-specific Tdtomato mice. Cardiac deletion of Gas6 gene mice was used to evaluate the impaired heart regeneration. CM-specific Gas6 overexpression mice were generated using adeno-associated virus system to evaluate Gas6 pro-proliferate and protect effect after ischemic injury. Secreted signal peptide truncation (ΔSP-Gas6) and predicted functional region truncation (ΔCC-Gas6) were generated to identify the intracellular mechanism of Gas6 via Sav1/p-mst1/p-lats1/ p-yap axis. Results We found that Gas6 expression is enriched in CM4 and the expression profile correlates with the expression of the CM4 marker Acta2, as well as cell-cycle genes Mki67 and Aurora B, in individual CM. Gas6 was highly expressed in the neonatal heart and continued to increase during the neonatal heart regenerative process, but was nearly absent in the adult heart under physiological and ischemia conditions. CM-specific Gas6 deletion resulted in significantly reduced CM proliferation rate and increased CM size in P0 hearts, and congestive heart failure in P28 hearts. Conversely, sustained CM-specific overexpression of Gas6 in postnatal hearts increased CM division, elevated CM numbers, and reduced CM size; and reduced infarct scar area and enhanced cardiac function after ischemia injury. Pre-overexpression of Gas6 in adult CMs protected adult cardiac function against ischemia injury, indicated by the activation of mitosis, angiogenesis and reduced oxidative phosphorylation. In primary neonatal CMs, both Gas6 and ΔSP-Gas6promoted CM proliferation with sarcomere disassembly. Mechanistically, Gas6 interacted with Sav1to inhibit Mst1recruitment and Mst1/Lats1/Yap phosphorylation cascade, therefore, promoted Yap translocation into nucleus. Conclusions We demonstrate that Gas6 is an important intrinsic regulator of neonatal heart regeneration, which acts as a promoter of Yap transcriptional activity to enhance CM proliferation. Supplementing Gas6 in the adult heart represents a potential therapeutic approach for cardiac repair.