ABSTRACT: Hair follicle (HF) regeneration begins when communication between quiescent epithelial stem cells (SCs) and underlying mesenchymal dermal papillae (DP) generates sufficient activating cues to overcome repressive BMP signals from surrounding niche cells. We uncovered a hitherto unrecognized DP transmitter, TGFβ2, which activates Smad2/3 transiently in HFSCs concomitant with entry into tissue regeneration. We used microarrays to detect the genes specifically affected by TGFß receptor II-deficient mice upon HFSC activation. Hair follicle stem cells (HFSCs) of hair gem (HG) and bulge, and total skin keratinocytes were FACS-purified from the mouse back skin at 2nd telogen-to-anagen transition stages.
Project description:Mouse hair follicles undergo synchronized cycles. Cyclical regeneration and hair growth is fueled by hair follicle stem cells (HFSCs). We used ChIP-seq to unfold genome-wide chromatin landscapes of Nfatc1 and dissect the biological relevence of its upstream BMP signaling in HFSC aging. Telogen quiescent hair follicle stem cells (HFSCs) were FACS-purified for ChIP-sequcencing.
Project description:Mouse hair follicles undergo synchronized cycles. Cyclical regeneration and hair growth is fueled by hair follicle stem cells (HFSCs). HFSCs regenerate hair in response to canonical Wnt signalling. We used RNA-seq to unfold genome-wide chromatin landscapes of β-catenin within the native HFSC-niche. Overall design: β-catenin control and cKO hair follicle stem cells (HFSCs) at the onset of anagen skins were FACS-purified for RNA-sequcencing. Telogen quiescent hair follicle stem cells (HFSCs) were FACS-purified for ChIP-sequcencing. Telogen>anagen activated bulge hair follicle stem cells (HFSCs) were FACS-purified for RNA-sequcencing.
Project description:Mouse hair follicles undergo synchronized cycles. Cyclical regeneration and hair growth is fueled by hair follicle stem cells (HFSCs). We used RNA-seq to identify SOX9-dependent transcriptional changes and ChIP-seq to identify SOX9-bound genes in HF-SCs. Telogen quiescent hair follicle stem cells (HFSCs) and intefollicular epidermal cells (IFE) were FACS-purified for ChIP-sequcencing and HFSCs for RNA-Sequencing
Project description:We report downstream gene expression changes in stem cells of the adult mouse hair follicle upon conditional ablating of the transcription factor Forkhead Box C1 transcription factor (FOXC1). Hair follicles undergo cycles of rest (telogen; Tel) and regeneration (anagen; Ana). As such, we performed our analysis on these two different stages of hair follicles. mRNA-sequencing of WT vs. Foxc1-conditional or inducible KO (Foxc1-cKO or iKO) hair follicle stem cells (HFSCs) purified from mouse dorsal back skin by flow-activated cell sorting (FACS).
Project description:Hair loss is one of the typical aging phenotypes in mammals, yet the underlying mechanism(s) is unclear. Here we report that hair follicle stem cell (HFSC) aging causes the stepwise miniaturization of hair follicles and eventual hair loss both in wild-type mice and in humans. In vivo fate analysis of HFSCs revealed that the DNA damage response in HFSCs causes proteolysis of Type XVII Collagen (COL17A1/BP180) to trigger “HFSC aging”, characterized by their loss of stemness signature and epidermal commitment. Those aged HFSCs are cyclically eliminated from the skin through their terminal epidermal differentiation, thereby causing hair follicle miniaturization. That process can be recapitulated by Col17a1 deficiency and prevented by forced maintenance of COL17A1 in HFSCs, demonstrating that stem cell homeostasis is the keystone against ultimate execution of the tissue/organ aging program. Overall design: Hair follicle stem cells from young, aged and Col17a1cKO mice were FACS isolated and RNA was extracted before being hybridized Agillent microarray. We obtained a gene of list by comparing Young and Agedand Col17a1 cKO cells.
Project description:In many organs, adult stem cells are uniquely poised to serve as cancer cells of origin. In the epidermis, hair follicle stem cells (HFSCs) cycle through stages of quiescence (telogen) and proliferation (anagen) to drive hair growth. Within the hair follicle, HFSCs are capable of initiating squamous cell carcinoma, yet it is unclear how the hair cycle contributes to tumorigenesis. The data presented here show that HFSCs are unable to initiate tumors during the quiescent phase of the hair cycle, indicating that the mechanisms that keep HFSCs dormant are dominant to gain of oncogenes (Ras) or loss of tumor suppressors (p53). Instead, prolonged oncogenic stimuli only exert their effects when HFSC quiescence mechanisms are removed by normal HFSC activation. Furthermore, Pten activity is necessary for quiescence based tumor suppression, since Pten deletion alleviates this stem cell specific ability without affecting proliferation per se. Small RNAs were cloned from Trizol-lysed cells sorted from mouse skin and sequenced with the Illumina HiSeq2000.
Project description:Purpose: The goals of this study were to identify Pathways that Tregs utilize in the skin to maintain tissue homesostasis in the context of hair follcile cycling. Methods: Hair follicle stem cells (HFSCs), in the presence and absence of Tregs, were induced into cycle through a model of depilation induced anagen. HFSCs were then purified by cell sorting at day 4 post depilation based on cell surface marker expression (CD45 negative, MHC Class II negative, Sca-1 negative, EpCAM negative, CD34 Positive, Integrin alpha6 High) to generate mRNA transcription profiles. Results: Transcriptional profiling revealed a significant reduction in proliferation/differentiation associated genes expressed in HFSCs isolated from Treg depleted mice Conclusion: Our study represents the first detailed analysis of Treg modulation of tissue stem cell behaviour mRNA profiles of HFSC isolated from 7-10 week old WT and Foxp3-DTR mice at 4 days post depilation of dorsal skin.
Project description:Recent developments in animal models (Morris et al., 2004; Tumbar et al., 2004) as well as the discovery of cell surface markers (Jones and Watt, 1993; Tani et al., 2000; Trempus et al., 2003; Nijhof et al., 2006) have made it possible to isolate living epidermal hair follicle stem cells (HFSCs) from mouse skin, facilitating the study of the biological and molecular features inherent to HFSCs. A complexity of stem and progenitor cell populations within the hair follicle has been revealed. Here, we report comprehensive profiling of mouse CD34-expressing HFSCs using the Agilent mouse oligo microarray platform in order to extend and enrich the existing HFSC databases. Experiment Overall Design: Total RNA was prepared from CD34(+) and CD34(-) keratinocytes obtained from three biological replicates, labeled with 2 different fluorescent dyes, and hybridized to the Agilent oligo-microarrays containing ~22,000 mouse genes and Expression Sequence Tag (EST) probes (supplemental methods). The experiment contained technical dye-flips of each pairwise comparison.
Project description:Mouse hair follicles undergo synchronized cycles. Cyclical regeneration and hair growth is fueled by hair follicle stem cells (HFSCs) and transit-amplifying cells (TACs). We used ChIP-seq to unfold genome-wide chromatin landscapes of H3K27ac and Med1 to identify super-enhancers and dissect their biological relevance in cell identity and plasticity of HFSCs in vivo and in vitro. Overall design: Telogen quiescent hair follicle stem cells (HFSCs) and transit-amplifying cells (TACs) were FACS-purified for ChIP-sequcencing. Furthermore, cultured HFSCs were used for ChIP-sequencing.
Project description:We performed RNA_Seq on purified hair follicle stem cells (HFSCs)and their direct progenty, transit amplifying cells (TACs) using temorally and spatially regulated Cre lines. Consequences of loss of Bmpr1a in either HFSC (K15-CrePGR;Bmpr1a fl/fl), or TACs (1. Shh-CreER:Bmpr1a fl/fl or 2. K15-CrePGR;Bmpr1a fl/fl derived)