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Integrin α3β1-dependent mRNA expression profile of hair bulge keratinocytes and their progeny during DMBA/TPA-mediated skin tumorigenesis

ABSTRACT: Integrin α3β1-dependent mRNA expression profile of hair bulge keratinocytes and their progeny during DMBA/TPA-mediated skin tumorigenesis

PROVIDER: PRJNA560882 | ENA |

REPOSITORIES: ENA

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Integrin α3β1-dependent mRNA expression profile of hair bulge keratinocytes and their progeny during DMBA/TPA-mediated skin tumorigenesis

Project description:Integrin α3β1 in hair bulge population affects the tumor formation following DMBA/TPA carcinogenesis protocol, however linage tracing showed that hair bulge cells do not largely contributre to tumor mass. To investigate whether α3β1 might affect tumor promoting environment and to better understand its role in hair bulge population during skin tumorigenesis, we determined the expresison profile of hair bulge-originating keratinocytes isolted from α3β1KO and WT mice during initiation of tumorigenesis.

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Profiling of genes expressed in cultured human keratinocytes established from the bulge region of hair follicles.

Project description:We established a culture method of human keratinocytes from the bulge region of a plucked hair follicle, that contains multipotent epithelial stem cells with high proliferative potential. Using our method, keratinocyte cultures were successfully obtained from all subjects without invasive skin biopsies. We compared the gene expression profiles between the cultured keratinocytes derived from human hair-follicle-bulge (bulge–derived keratinocytes; BDKs) and neonatal human epidermal keratinocytes (NHEKs), and between BDKs from donors with atopic dermatitis and non-atopic controls using microarray analysis. Keywords: expressin profiling

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Expression data from squamous cell carcinoma stem cells, epidermal progenitor cells and hair follicle bulge stem cells

Project description:We used microarrays to assess the global gene expression profiles of cancer stem cells which were isolated from cutaneous squamous cell carcinomas which developed when WT, TGF beta receptor II ko, FAK KO, and TGF beta receptor II/FAK double KO were subjected to continuous DMBA treatment. Squamous cell carcinoma stem cells were compared to epidermal progenitor cells (CD49fhighCD34low) and hair follicle bulge stem cells (CD49fhighCD34high). Squamous cell carcinoma was initiated by continuous DMBA treatment (complete carcinogenesis). Cancer cells were sorted based on high CD29 and CD49f expression, infected with a retrovirus expressing a triple modality reporter (luciferase, RFP, thymidine kinase) and grafted onto nude recipient mice. RFP positive lineage was separated into CD29 high, CD49f high and CD34 low; or CD29high, CD49f high and CD34high squamous cell carcinoma initiating cells.

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Project description:In most tissues, stem cells (SCs) comprise a heterogeneous cell population. Previous reports showed that slow cycling label-retaining cells (LRCs) are present in two prominent hair follicle SC populations, basal and suprabasal bulge SCs (bBSCs, sbBSCs). CD34+/itga6low sbBSC and neighbouring CD34+/itga6high bBSCs seem similar with both SC populations able to self-renew in vitro and to generate new hair follicles when grafted. Until now, functional differences have not been reported and molecular mechanisms underlying a positional encoded, intrinsic bulge SC heterogeneity and the relations to SC plasticity and tissue function are not well understood. Following fluorescence activated cell sorting for CD34 and integrin alpha6 of keratinocytes from 7 weeks old mice, we provide sequencing data for two prominent hair follicle SC populations, bBSC and sbBSC, suggesting distinct and overlapping functions in skin homeostasis.

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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

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Profiling of genes expressed in cultured human keratinocytes established from the bulge region of hair follicles.

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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.

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