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Blood endothelial Alk1-BMP4 signaling axis ensures adult hair follicle stem cell timely activation

ABSTRACT: To investigate the gene expression changes in the hair follicle stem cells (bulge cells, HFSCs) after perturbing the vascular niche by knocking out Alk1 from the endothelial cells using Cdh5CreERT2 driver

ORGANISM(S): Mus musculus

PROVIDER: GSE223723 | GEO | 2023/02/28

REPOSITORIES: GEO

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Blood endothelial ALK1-BMP4 signaling axis regulates adult hair follicle stem cell activation

Project description: Not available

| S-SCDT-10_15252-EMBJ_2022112196 | biostudies-other

"Alk1 acts in non-endothelial VE-cadherin+ perineurial cells to maintain nerve branching during hair homeostasis" and "Blood endothelial ALK1-BMP4 signaling axis regulates adult hair follicle stem cell activation"

Project description:Purpose: To comprehensively characterize VE-cadherin expressing lineages during adult skin homeostasis, to dissect their population-specific transcriptomic heterogeneity, and to investigate role of Alk1 in these linegease in vivo. Methods: We performed 10x genomics single-cell RNA sequencing (scRNA-seq) of FACS sorted tdTomato+ cells from tdTomato;Cdh5-CreERT2;Krt14-H2BGFP mice (for WT) or Alk1 flox flox;tdTomato;Cdh5-CreERT2 (For Alk1KO) back skin at indicated stage/genotype. FACS purified tdTomato+ single-cell suspension was processed for the barcoded single-cell 3′ cDNA libraries generation using Chromium Single Cell 3′ gel bead and library Kit v3. The final libraries were quantified using Agilent Bioanalyzer high sensitivity DNA chip and sequenced using an Illumina NextSeq-500. The raw data files were demultiplexed to generate the sample-specific FASTQ files, which were aligned to the mouse reference genome (mm10-2020-A) using the 10x Genomics Cell Ranger pipeline (v6.0.1). The raw scRNA-seq data was processed using Cell Ranger from the 10x platform to generate expression matrix for each sample that was further analyzed in R using the Seurat package version 4.0. Only high-quality cells that had between 200 and 5000 genes expressed and had under 10% of the UMIs mapped to mitochondrial genes were retained for further analysis. Results: We obtained more than 2000 high quality cells at each stage/genotype for further analysis by Seurat pipeline Conclusions: Obtained high quality single cell transcriptomic data to dissect cellular and molecular heterogeneity of VE-cadherin expressing endothelial and non-endothelial lineages in adult mouse back skin.

2023-03-01 | GSE211381 | GEO

Paracrine TGFß Signaling Counterbalances BMP-Mediated Repression in Hair Follicle Stem Cell Activation

Project description: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.

2012-01-04 | E-GEOD-33471 | biostudies-arrayexpress

Nfatc1 Orchestrates Aging in Hair Follicle Stem Cells

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.

2013-11-14 | E-GEOD-52328 | biostudies-arrayexpress

Characterization and isolation of stem cell enriched human hair follicle bulge cells

Project description:The human hair follicle bulge is an important niche for keratinocyte stem cells (KSC). Elucidation of human bulge cell biology could be facilitated by analysis of global gene expression profiles and identification of unique cell surface markers. The lack of distinctive bulge morphology in human hair follicles has hampered studies of bulge cells and KSC. In this study, we determined the distribution of label-retaining cells to carefully define the human anagen bulge. Using navigated-laser capture microdissection, bulge cells and outer root sheath cells from other follicle regions were obtained and analyzed with cDNA microarrays. Gene transcripts encoding inhibitors of WNT and Activin/BMP signaling were over-represented in the bulge while genes responsible for cell proliferation were under-represented, consistent with quiescent non-cycling KSC in anagen follicles. Positive markers for bulge cells included CD200, PHLDA1, follistatin, and frizzled homolog 1 while CD24, 34, 71 and 146 were preferentially expressed by non-bulge keratinocytes. Importantly, CD200+ cells (CD200hi24lo34lo71lo146lo) obtained from hair follicle suspensions demonstrated high colony forming efficiency in clonogenic assays, indicating successful enrichment of living human bulge stem cells. Keywords: Affymetrix micrarray analysis of human hair follicles

2005-12-06 | GSE3419 | GEO

Architectural niche organization by LHX2 is linked to hair follicle stem cell function

Project description:To identify direct LHX2 target genes in HFSCs, we performed chromatin immunoprecipitation and deep sequencing (ChIP-seq) analysis using FACS-isolated HFSCs. Three independent LHX2 ChIP-seq experiments were conducted.

2013-09-05 | E-GEOD-48068 | biostudies-arrayexpress

Transcription profiling of human stem cells enriched for hair follicle bulge cells

Project description:The human hair follicle bulge is an important niche for keratinocyte stem cells (KSCs). Elucidation of human bulge cell biology could be facilitated by analysis of global gene expression profiles and identification of unique cell-surface markers. The lack of distinctive bulge morphology in human hair follicles has hampered studies of bulge cells and KSCs. In this study, we determined the distribution of label-retaining cells to define the human anagen bulge. Using navigated laser capture microdissection, bulge cells and outer root sheath cells from other follicle regions were obtained and analyzed with cDNA microarrays. Gene transcripts encoding inhibitors of WNT and activin/bone morphogenic protein signaling were overrepresented in the bulge, while genes responsible for cell proliferation were underrepresented, consistent with the existence of quiescent noncycling KSCs in anagen follicles. Positive markers for bulge cells included CD200, PHLDA1, follistatin, and frizzled homolog 1, while CD24, CD34, CD71, and CD146 were preferentially expressed by non-bulge keratinocytes. Importantly, CD200+ cells (CD200hiCD24loCD34loCD71loCD146lo) obtained from hair follicle suspensions demonstrated high colony-forming efficiency in clonogenic assays, indicating successful enrichment of living human bulge stem cells. The stem cell behavior of enriched bulge cells and their utility for gene therapy and hair regeneration will need to be assessed in in vivo assays.

2007-11-08 | E-GEOD-3419 | biostudies-arrayexpress

Architectural niche organization by LHX2 is linked to hair follicle stem cell function

Project description:To identify direct LHX2 target genes in HFSCs, we performed chromatin immunoprecipitation and deep sequencing (ChIP-seq) analysis using FACS-isolated HFSCs.

2013-09-05 | GSE48068 | GEO

Signaling couples hair follicle stem cell quiescence with reduced histone H3K4/K9/K27me3 for proper tissue homeostasis

Project description:Mechanisms of plasticity to acquire different cell fates are critical for adult stem cell (SC) potential, yet are poorly understood. Reduced global histone methylation is an epigenetic state known to mediate plasticity in cultured embryonic SCs and T cell progenitors. We used mouse hair follicle stem cells (HFSCs) at two different hair cycle stages (early anagen and late catagen) to compare the genome-wide changes in the levels of histone modification marks H3K4me3, H3K9me3, and H3K27me3. Hair follicle stem cells from Early Anagen (EA-HFSCs) and Late Catagen (LC-HFSCs), and their non-HFSCs counterparts (nEA-HFSCs and nLC-HFSCs), were FACS-isolated for Chromatin Immunoprecipitation followed by sequencing (ChIP-seq) analysis of H3K4me3, H3K9me3, and H3K27me3.

2016-04-13 | E-GEOD-78749 | biostudies-arrayexpress

RNA sequencing of ILK-deficient hair follicle bulge stem cells

Project description:We sequenced mRNA from FACS purified hair follicle bulge stem cells from 21 d old control and ILK-deficient mice, 3 biological replicates each Examination of mRNA levels in control and ILK-deficient hair follicle bulge stem cells

2015-09-01 | E-GEOD-68846 | biostudies-arrayexpress

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