RESUMO
Although long noncoding RNAs (lncRNAs) are transcripts that do not encode proteins by definition, some lncRNAs actually contain small open reading frames that are translated. TINCR (terminal differentiation-induced ncRNA) has been recognized as a lncRNA that contributes to keratinocyte differentiation. However, we here show that TINCR encodes a ubiquitin-like protein that is well conserved among species and whose expression was confirmed by the generation of mice harboring a FLAG epitope tag sequence in the endogenous open reading frame as well as by targeted proteomics. Forced expression of this protein promoted cell cycle progression in normal human epidermal keratinocytes, and mice lacking this protein manifested a delay in skin wound healing associated with attenuated cell cycle progression in keratinocytes. We termed this protein TINCR-encoded ubiquitin-like protein (TUBL), and our results reveal a role for TINCR in the regulation of keratinocyte proliferation and skin regeneration that is dependent on TUBL.
Assuntos
Queratinócitos/citologia , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Animais , Ciclo Celular , Diferenciação Celular , Células Cultivadas , Regulação da Expressão Gênica , Técnicas de Introdução de Genes , Humanos , Queratinócitos/metabolismo , Camundongos , Fases de Leitura Aberta , Proteômica , Ubiquitinas/genética , Ubiquitinas/metabolismo , CicatrizaçãoRESUMO
BACKGROUND: The circadian rhythm controls multiple biological processes, including immune responses; however, its impact on cutaneous adaptive immune response remains unclear. METHODS: We used a well-established cutaneous type IV allergy model, contact hypersensitivity (CHS). We induced CHS using dinitrofluorobenzene (DNFB). Mice were sensitized and elicited with DNFB in the daytime or at night. RESULTS: In mice, a nocturnally active animal, we found that ear swelling increased when mice were sensitized at night compared with in the daytime. In addition, cell proliferation and cytokine production in the draining lymph nodes (LNs) were promoted when sensitized at night. We hypothesized that these differences were due to the oscillation of leukocyte distribution in the body through the circadian production of adrenergic hormones. Administration of a ß2-adrenergic receptor (ß2AR) agonist salbutamol in the daytime decreased the number of immune cells in blood and increased the number of immune cells in LNs. In contrast, a ß2AR antagonist ICI18551 administration at night increased the number of immune cells in blood and decreased the number of immune cells in LNs. Accordingly, the severity of CHS response was exacerbated by salbutamol administration in the daytime and attenuated by ICI18551 administration at night. CONCLUSION: Our study demonstrated that the magnitude of adaptive CHS response depends on the circadian rhythm and this knowledge may improve the management of allergic contact dermatitis (ACD) in humans.
Assuntos
Ritmo Circadiano , Dermatite Alérgica de Contato , Albuterol , Animais , Dinitrofluorbenzeno , Humanos , Camundongos , Camundongos Endogâmicos BALB C , PeleAssuntos
Dermatite/imunologia , Interleucina-17/metabolismo , Psoríase/imunologia , Linfócitos T/imunologia , Tromboxano A2/metabolismo , Animais , Modelos Animais de Doenças , Humanos , Imiquimode , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Receptores de Tromboxano A2 e Prostaglandina H2/genética , Receptores de Tromboxano A2 e Prostaglandina H2/metabolismo , Tromboxano-A Sintase/genética , Tromboxano-A Sintase/metabolismoRESUMO
Vesiculobullous dermatomyositis (VD) is a rare manifestation of dermatomyositis (DM) and has been suggested to be associated with malignancy. Although the myositis-specific autoantibodies are associated with distinct clinical presentations of DM, those associated with VD remain unclear. Here, we present the case of a 54-year-old man with VD who tested positive for anti-nuclear matrix protein 2 (NXP-2) antibody, one of the DM-specific autoantibodies. Serological and histopathological findings did not support autoimmune blistering disease. Physical and histological findings suggested that the severe edema in combination with the interface dermatitis of DM contributed to blister formation. Although a systemic examination was performed, no evidence of malignancy was found. Following initiation of immunosuppressive therapy, the patient showed significant improvement in both skin lesions and myositis. This case represents the first report of anti-NXP-2-positive VD without malignancy or autoimmune blistering disease. Subcutaneous edema, a characteristic feature of anti-NXP-2-positive DM, could be related to the formation of VD.
Assuntos
Infecções por Mycobacterium/microbiologia , Mycobacterium/isolamento & purificação , Dermatopatias Bacterianas/microbiologia , Idoso , Humanos , Masculino , Infecções por Mycobacterium/tratamento farmacológico , Dermatopatias Bacterianas/tratamento farmacológico , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Sarcoidosis is a disease of unknown etiology in which granulomas form throughout the body and is typically treated with glucocorticoids, but there are no approved steroid-sparing alternatives. Here, we investigated the mechanism of granuloma formation using single-cell RNA-Seq in sarcoidosis patients. We observed that the percentages of triggering receptor expressed on myeloid cells 2-positive (TREM2-positive) macrophages expressing angiotensin-converting enzyme (ACE) and lysozyme, diagnostic makers of sarcoidosis, were increased in cutaneous sarcoidosis granulomas. Macrophages in the sarcoidosis lesion were hypermetabolic, especially in the pentose phosphate pathway (PPP). Expression of the PPP enzymes, such as fructose-1,6-bisphosphatase 1 (FBP1), was elevated in both systemic granuloma lesions and serum of sarcoidosis patients. Granuloma formation was attenuated by the PPP inhibitors in in vitro giant cell and in vivo murine granuloma models. These results suggest that the PPP may be a promising target for developing therapeutics for sarcoidosis.
Assuntos
Via de Pentose Fosfato , Sarcoidose , Humanos , Animais , Camundongos , Sarcoidose/diagnóstico , Sarcoidose/etiologia , Sarcoidose/patologia , Granuloma , Macrófagos/patologia , GlucocorticoidesRESUMO
The skin barrier protects our body from external insults and water loss through a specialized layer, the stratum corneum. The stratum corneum, an accumulation of dead keratinocytes (corneocytes), comprises lipids and supporting cell bodies. We propose a framework of lipid-filled polymer sheet of corneocytes, a unique structure that achieves flexibility and robustness, updating the rigid image of the historical bricks-and-mortar model. The polymerization of polymer sheet (cornification) by cell death of keratinocytes (corneoptosis) is delicately and dynamically controlled by cytoplasmic calcium ion and pH. Understanding the structure and formation of the stratum corneum can lead to better treatments for skin diseases and a better understanding of the evolution of the stratum corneum.
Assuntos
Epiderme , Polímeros , Diferenciação Celular , Epiderme/metabolismo , Humanos , Queratinócitos/metabolismo , Polímeros/metabolismo , PeleRESUMO
Stem cell loss causes tissue deterioration associated with aging. The accumulation of genomic and oxidative stress-induced DNA damage is an intrinsic cue for stem cell loss1,2; however, whether there is an external microenvironmental cue that triggers stem cell loss remains unclear. Here we report that the involution of skin vasculature causes dermal stiffening that augments the differentiation and hemidesmosome fragility of interfollicular epidermal stem cells (IFESCs) in aged mouse skin. Aging-related IFESC dysregulation occurs in plantar and tail skin, and is correlated with prolonged calcium influx, which is contributed by the mechanoresponsive ion channel Piezo1 (ref. 3). Epidermal deletion of Piezo1 ameliorated IFESC dysregulation in aged skin, whereas Piezo1 activation augmented IFESC differentiation and hemidesmosome fragility in young mice. The dermis stiffened with age, which was accompanied by dermal vasculature atrophy. Conversely, induction of the dermal vasculature softened the dermis and ameliorated IFESC dysregulation in aged skin. Single-cell RNA sequencing of dermal fibroblasts identified an aging-associated anti-angiogenetic secretory molecule, pentraxin 3 (ref. 4), which caused dermal sclerotization and IFESC dysregulation in aged skin. Our findings show that the vasculature softens the microenvironment for stem cell maintenance and provide a potential mechanobiology-based therapeutic strategy against skin disorders in aging.
Assuntos
Epiderme , Pele , Camundongos , Animais , Epiderme/fisiologia , Diferenciação Celular/genética , Células-Tronco , Atrofia/patologia , Canais Iônicos/genéticaRESUMO
Surfactant-induced cumulative irritant contact dermatitis (ICD) is a common and clinically important skin disorder. CCL2 is known to mediate inflammation after tissue damage in various organs. Thus, we investigated whether and how CCL2 contributes to the development of murine cumulative ICD induced by a common surfactant, SDS. Wild-type mice treated topically with SDS for 6 consecutive days developed skin inflammation that recapitulated the features of human cumulative ICD, including barrier disruption, epidermal thickening, and neutrophil accumulation. CCL2 was upregulated in SDS-treated skin, and local CCL2 blockade attenuated SDS-induced ICD. SDS-induced ICD and neutrophil accumulation were also attenuated in mice deficient in CCR2, the receptor for CCL2. Neutrophil depletion alleviated SDS-induced ICD, suggesting that impaired neutrophil accumulation was responsible for the amelioration of ICD in CCR2-deficient mice. In RNA-sequencing analyses of SDS-treated skin, the expression levels of Il1b in Ccr2-deficient mice were highly downregulated compared with those in wild-type mice. Furthermore, the intradermal administration of IL-1ß in the SDS-treated skin of CCR2-deficient mice restored the local accumulation of neutrophils and the development of ICD. Collectively, our results suggest that CCL2âCCR2 signaling in the skin critically promotes the development of SDS-induced ICD by inducing IL-1ß expression for neutrophil accumulation.
Assuntos
Dermatite Irritante , Neutrófilos , Animais , Quimiocina CCL2 , Dermatite Irritante/metabolismo , Inflamação/metabolismo , Interleucina-1beta , Irritantes/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neutrófilos/metabolismo , Receptores CCR2/genética , Receptores CCR2/metabolismo , Receptores de Quimiocinas/metabolismo , Pele/metabolismo , TensoativosRESUMO
The geometric organization of collagen fibers in human reticular dermis and its relationship to that of elastic fibers remain unclear. The tight packing and complex intertwining of dermal collagen fibers hinder accurate analysis of fiber orientation. We hypothesized that combined multiphoton microscopy and biaxial extension could overcome this issue. Continuous observation of fresh dermal sheets under biaxial extension revealed that the geometry of the elastic fiber network is maintained during expansion. Full-thickness human thigh skin samples were biaxially extended and cleared to visualize the entire reticular dermis. Throughout the dermis, collagen fibers straightened with increased inter-fiber spaces, making them more clearly identifiable after extension. The distribution of collagen fibers was evaluated with compilation of local orientation data. Two or three modes were confirmed in all superficial reticular layer samples. A high degree of local similarities in the direction of collagen and elastic fibers was observed. More than 80% of fibers had directional differences of ≤15°, regardless of layer. Understanding the geometric organization of fibers in the reticular dermis improves the understanding of mechanisms underlying the pliability of human skin. Combined multiphoton imaging and biaxial extension provides a research tool for studying the fibrous microarchitecture of the skin.
Assuntos
Colágeno/análise , Derme/diagnóstico por imagem , Tecido Elástico/diagnóstico por imagem , Microscopia de Fluorescência por Excitação Multifotônica/métodos , Reticulina/análise , Adulto , Idoso , Derme/química , Tecido Elástico/química , Elastina/análise , Feminino , Fibrilinas/análise , Análise de Fourier , Voluntários Saudáveis , Humanos , Ligamentos , Masculino , Microfibrilas , Pessoa de Meia-Idade , Doadores de TecidosAssuntos
Infecções por Citomegalovirus/virologia , Citomegalovirus/patogenicidade , Dermatite Esfoliativa/etiologia , Doença Enxerto-Hospedeiro/etiologia , Simplexvirus/patogenicidade , Estomatite Herpética/virologia , Timoma/complicações , Neoplasias do Timo/complicações , Ativação Viral , Antivirais/uso terapêutico , Biópsia , Infecções por Citomegalovirus/diagnóstico , Infecções por Citomegalovirus/tratamento farmacológico , Dermatite Esfoliativa/diagnóstico , Dermatite Esfoliativa/terapia , Feminino , Glucocorticoides/uso terapêutico , Doença Enxerto-Hospedeiro/diagnóstico , Doença Enxerto-Hospedeiro/terapia , Humanos , Pessoa de Meia-Idade , Recidiva , Estomatite Herpética/diagnóstico , Estomatite Herpética/tratamento farmacológico , Timoma/secundário , Neoplasias do Timo/patologia , Resultado do Tratamento , Terapia UltravioletaRESUMO
Epidermal keratinocytes achieve sequential differentiation from basal to granular layers, and undergo a specific programmed cell death, cornification, to form an indispensable barrier of the body. Although elevation of the cytoplasmic calcium ion concentration ([Ca2+]i) is one of the factors predicted to regulate cornification, the dynamics of [Ca2+]i in epidermal keratinocytes is largely unknown. Here using intravital imaging, we captured the dynamics of [Ca2+]i in mouse skin. [Ca2+]i was elevated in basal cells on the second time scale in three spatiotemporally distinct patterns. The transient elevation of [Ca2+]i also occurred at the most apical granular layer at a single cell level, and lasted for approximately 40 min. The transient elevation of [Ca2+]i at the granular layer was followed by cornification, which was completed within 10 min. This study demonstrates the tightly regulated elevation of [Ca2+]i preceding the cornification of epidermal keratinocytes, providing possible clues to the mechanisms of cornification.
Assuntos
Cálcio/metabolismo , Diferenciação Celular , Células Epidérmicas/citologia , Células Epidérmicas/metabolismo , Íons/metabolismo , Queratinócitos/citologia , Queratinócitos/metabolismo , Animais , Forma Celular , Células Cultivadas , Citoplasma/metabolismo , Imunofluorescência , Expressão Gênica , Genes Reporter , Camundongos , Análise de Célula ÚnicaAssuntos
Hiperplasia do Linfonodo Gigante , Síndromes de Imunodeficiência , Transtornos Linfoproliferativos , Síndromes Paraneoplásicas , Pênfigo , Humanos , Doença Iatrogênica , Síndromes de Imunodeficiência/complicações , Transtornos Linfoproliferativos/diagnóstico , Transtornos Linfoproliferativos/etiologia , Síndromes Paraneoplásicas/complicações , Síndromes Paraneoplásicas/etiologia , Pênfigo/complicações , Pênfigo/etiologiaRESUMO
Resolvin E1 (RvE1) is a lipid mediator derived from ω3 polyunsaturated fatty acids that exerts potent antiinflammatory roles in several murine models. The antiinflammatory mechanism of RvE1 in acquired immune responses has been attributed to attenuation of cytokine production by dendritic cells (DCs). In this study, we newly investigated the effect of RvE1 on DC motility using two-photon microscopy in a contact hypersensitivity (CHS) model and found that RvE1 impaired DC motility in the skin. In addition, RvE1 attenuated T cell priming in the draining lymph nodes and effector T cell activation in the skin, which led to the reduced skin inflammation in CHS. In contrast, leukotriene B4 (LTB4) induced actin filament reorganization in DCs and increased DC motility by activating Cdc42 and Rac1 via BLT1, which was abrogated by RvE1. Collectively, our results suggest that RvE1 attenuates cutaneous acquired immune responses by inhibiting cutaneous DC motility, possibly through LTB4-BLT1 signaling blockade.