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Expression data from in vitro keratinocytes exposed to extracellular vesicles from UVA-irradiated melanocytes

ABSTRACT: Expression data from in vitro keratinocytes exposed to extracellular vesicles from UVA-irradiated melanocytes

PROVIDER: PRJNA601428 | ENA |

REPOSITORIES: ENA

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Expression data from in vitro keratinocytes exposed to extracellular vesicles from UVA-irradiated melanocytes

Project description:Analysis of altered gene expression in keratinocytes exposed to extracellular vesicles. In this dataset we include the respective expression data.

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Effect of enhanced melanin synthesis on the production of extracellular vesicles in melanocytes and communication with keratinocytes

Project description:Extracellular vesicle(EV) secretion and capture is an important method of communication between cells. Melanocytes and their surrounding keratinocytes form epidermal melanin units in the thin, outermost layer of the skin to protect the skin from ultraviolet radiation damage. It is important to understand the mechanisms by which changes in melanocyte EV signal transduction influence pigmentation. Mouse melanocytes (melan-α cell) overexpressing MITF and CD63-GFP were used to to investigate the effect on vesicle formation by WB and SEM. Coculturing of melanocyte with keratinocyte(PAM212) was used to explore the polarization of vesicles and transfer between cells by immunofluorescence, flow cytometry. EVs from melanocytes were extracted and used to investigate changes in the miRNA profile of EVs, and the effect of exosomal miR-28 on keratinocyte growth and transcriptomics was analyzed by flow cytometry, TEM, particle size analysis, WB, the iCELLigence system and RNA-seq analysis.Our results indicate that melanocytes in the skin affected vesicle synthesis under the condition of enhanced melanin synthesis function: the expression of CD81, VPS37, VPS36B and Rab27b (melanosome and vesicle transport protein during ESCRT machinery) increased. Keratinocytes can cause the polarization of vesicles in melanocytes and capture EVs from melanocytes via plasmalemma fusion. Analysis of the miRNA profile derived from melanocyte EVs showed that the expression levels of miR-125, miR-130, miR-211, miR-127, miR-692 and miR-709 were increased, while the expression levels of miR-28, miR-744, let-7g and miR-6240 were decreased. The content of miR-28 in EVs may be reduced to induce keratinocytes to prevent carcinogenesis, resist infection and prevent premature apoptosis.

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Exosomal miRNA sequencing of UVB-irradiated melanocytes and non-irradiated melanocytes

Project description:Exosomal miRNA sequencing of UVB-irradiated melanocytes and non-irradiated melanocytes

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Next Generation Sequencing Facilitated Differential Transcriptome Study of melanocytes treated with UVB radiation compared to matched fibroblasts and keratinocytes

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Solar UVR and exosomal miRNAs effect pre-mature senescence in melanocytes [RT-PCR]

Project description:The selection of specific miRNAs by exosomes and their release from cultured melanocytes after exposure to solar UVR (UVA+UVB) have activities in inducing these cells into a premature senescence. In this analysis, human TaqMan microRNA array cards A+B were used to measure the relative expression of miRNAs in exosomes isolated from the irradiated and un-irradiated melanocytes.

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Effect of enhanced melanin synthesis on the production of extracellular vesicles in melanocytes and communication with keratinocytes

Project description:Effect of enhanced melanin synthesis on the production of extracellular vesicles in melanocytes and communication with keratinocytes

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UV effects in mouse melanocytes

Project description:Ultraviolet (UV) radiation is a major melanoma risk factor, yet underlying mechanisms remain poorly understood. Here we introduce a mouse model permitting fluorescence-aided melanocyte imaging and isolation following in vivo UV irradiation. We use expression profiling to show that activated neonatal skin melanocytes isolated following a melanomagenic UVB dose bear a distinct, persistent interferon-response signature, including genes associated with immunoevasion. UVB-induced melanocyte activation, characterized by aberrant growth and migration, was abolished by antibody-mediated systemic blockade of interferon-gamma (IFN-gamma), but not type-I interferons. IFN-gamma was produced by macrophages recruited to neonatal skin by UVB-induced chemokine receptor Ccr2 ligands. Admixed recruited skin macrophages enhanced transplanted melanoma growth by inhibiting apoptosis; notably, IFN-gamma blockade abolished macrophage-associated melanoma growth and survival. IFN-gamma-producing macrophages were identified in 70% of human melanomas examined. Our data reveal an unanticipated role for IFN-gamma in promoting melanocytic cell survival/immunoevasion, and suggest IFN-gamma-R signaling represents a novel therapeutic melanoma target. Biologic replicates of UVA- and UVB-treated mouse melanocytes, as well as untreated mouse melanocytes and mouse keratinocytes, were used to define melanocyte expression signatures associated with UV treatment.

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Extracellular vesicles are transferred from melanocytes to keratinocytes after UVA irradiation.

Project description:Ultraviolet (UV) irradiation induces skin pigmentation, which relies on the intercellular crosstalk of melanin between melanocytes to keratinocytes. However, studying the separate effects of UVA and UVB irradiation reveals differences in cellular response. Herein, we show an immediate shedding of extracellular vesicles (EVs) from the plasma membrane when exposing human melanocytes to UVA, but not UVB. The EV-shedding is preceded by UVA-induced plasma membrane damage, which is rapidly repaired by Ca(2+)-dependent lysosomal exocytosis. Using co-cultures of melanocytes and keratinocytes, we show that EVs are preferably endocytosed by keratinocytes. Importantly, EV-formation is prevented by the inhibition of exocytosis and increased lysosomal pH but is not affected by actin and microtubule inhibitors. Melanosome transfer from melanocytes to keratinocytes is equally stimulated by UVA and UVB and depends on a functional cytoskeleton. In conclusion, we show a novel cell response after UVA irradiation, resulting in transfer of lysosome-derived EVs from melanocytes to keratinocytes.

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ROS stress responses of primary human epidermal keratinocytes

Project description:Primary human epidermal keratinocytes were exposed to in-vitro UVA-oxidized 1-palmitoyl-2-arachidonoyl-phosphatidylcholine or to UVA in presence and absence of a commercial UVA filter.

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RNASeq of juvenile keratinocytes, melanocytes, fibroblasts and whole skin

Project description:We used whole transcriptome sequencing (RNA-Seq) to assess differential gene expression of normal human melanocytes with respect to keratinocytes, fibroblasts and whole skin. We have identified a number of genes and pathways, which are characteristic or unique for MC compared to KC and FB. We also demonstrated the extent of the difference between genes detectable in MC culture and the whole skin. The present results reveal melanocytes as highly proliferative cells with stem cell-like properties.

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