RESUMEN
It is well established that different sites in healthy human skin are colonized by distinct microbial communities due to different physiological conditions. However, few studies have explored microbial heterogeneity between skin sites in diseased skin, such as atopic dermatitis (AD) lesions. To address this issue, we carried out deep analysis of the microbiome and transcriptome in the skin of a large cohort of AD patients and healthy volunteers, comparing two physiologically different sites: upper back and posterior thigh. Microbiome samples and biopsies were obtained from both lesional and nonlesional skin to identify changes related to the disease process. Transcriptome analysis revealed distinct disease-related gene expression profiles depending on anatomical location, with keratinization dominating the transcriptomic signatures in posterior thigh, and lipid metabolism in the upper back. Moreover, we show that relative abundance of Staphylococcus aureus is associated with disease severity in the posterior thigh, but not in the upper back. Our results suggest that AD may select for similar microbes in different anatomical locations-an "AD-like microbiome," but distinct microbial dynamics can still be observed when comparing posterior thigh to upper back. This study highlights the importance of considering the variability across skin sites when studying the development of skin inflammation.
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Dermatitis Atópica , Eccema , Microbiota , Dermatitis Atópica/genética , Humanos , Piel , Staphylococcus aureus/genéticaRESUMEN
Netherton syndrome (NS) is a rare, life-threatening syndrome caused by serine protease inhibitor Kazal-type 5 gene (SPINK 5) mutations, resulting in skin barrier defect, bacterial skin infections, and allergic sensitization in early childhood. Recent data on adult patients with NS suggest that the presence of Staphylococcus aureus further promotes barrier disruption and skin inflammation. We analyzed the skin microbiota by shotgun sequencing in 12 patients with NS from eight Finnish families with healthy family controls as the reference and correlated the findings with allergen-specific IgE prevalence, immune cell phenotype, and infection history of the patients. Compared with healthy family controls, skin microbiome diversity and normal skin site variability were measurably decreased in patients with NS. No correlation was found between allergic sensitization and skin microbiota as such, but low circulating CD57+ and/or CD8+ T cells significantly correlated with lower microbial diversity and less abundance of S. aureus (P < 0.05). S. aureus was the most prevalent species in patients with NS but also Streptococcus agalactiae was abundant in four patients. The genomic DNA relative abundance of S. aureus secreted virulence peptides and proteases PSMα, staphopain A, and staphopain B were increased in most of the samples of patients with NS, and their abundance was significantly (P < 0.05) associated with recurrent childhood skin infections, confirming the clinical relevance of S. aureus dominance in the NS skin microbiome.
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Adult neural stem cells, located in discrete brain regions, generate new neurons throughout life. These stem cells are specialized astrocytes, but astrocytes in other brain regions do not generate neurons under physiological conditions. After stroke, however, striatal astrocytes undergo neurogenesis in mice, triggered by decreased Notch signaling. We used single-cell RNA sequencing to characterize neurogenesis by Notch-depleted striatal astrocytes in vivo. Striatal astrocytes were located upstream of neural stem cells in the neuronal lineage. As astrocytes initiated neurogenesis, they became transcriptionally very similar to subventricular zone stem cells, progressing through a near-identical neurogenic program. Surprisingly, in the non-neurogenic cortex, Notch-depleted astrocytes also initiated neurogenesis. Yet, these cortical astrocytes, and many striatal ones, stalled before entering transit-amplifying divisions. Infusion of epidermal growth factor enabled stalled striatal astrocytes to resume neurogenesis. We conclude that parenchymal astrocytes are latent neural stem cells and that targeted interventions can guide them through their neuronal differentiation.
Regenerative medicine aims to help the body replace damaged or worn-out tissues, often by kick-starting its own intrinsic repair mechanisms. However, the brain cannot easily repair itself, and therefore poses a much greater challenge. This is because nerve cells or neurons, which underpin learning, memory, and many other abilities, are also the brain's greatest weakness when it comes to tissue repair. In most parts of the adult brain, neurons are never replaced after they die. This means that damage to brain tissue for example, after a stroke can have severe and long-lasting consequences. Neural stem cells are one type of brain cell that can turn into new neurons if needed, but they are only found in a few parts of the brain and cannot fix damage elsewhere. More recent work in mice has shown that astrocytes, a common type of support cell in the brain that help keep neurons healthy, could also generate new neurons following a stroke. However, the ability was restricted to small numbers of astrocytes in a specific part of the brain. Here, Magnusson et al. set out to determine the molecular mechanisms behind this regenerative process and why it is unique to certain astrocytes. The researchers used a technique called single-cell RNA sequencing to analyze the genetic activity within individual mouse astrocytes that had been exposed to conditions mimicking a stroke. This method revealed which genes are switched on or off, thus generating a profile of gene activity for each astrocyte analyzed. This experiment showed that the profiles of astrocytes that had started to produce neurons were in fact nearly identical to neural stem cells. Even the astrocytes that could not generate neurons took the first few steps towards this genetic state; however, they stalled early in the process. Treating the brains of mice withepidermal growth factor, a powerful molecular signal that stimulates cell growth, kick-started nerve cell production in a subset of these cells showing that at least some of the non-regenerative astrocytes could be stimulated to make neurons if given the right treatment. The results of this study shed new light on how some astrocytes in the brain gain the ability to form new neurons. In the future, this knowledge could help identify a source of replacement cells to regenerate the injured brain.
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Astrocitos , Células-Madre Neurales , Neurogénesis/genética , Transcriptoma/genética , Animales , Astrocitos/citología , Astrocitos/metabolismo , Células Cultivadas , Cuerpo Estriado/citología , Cuerpo Estriado/metabolismo , Factor de Crecimiento Epidérmico/metabolismo , Ratones , Células-Madre Neurales/citología , Células-Madre Neurales/metabolismo , ARN/análisis , ARN/genética , ARN/metabolismoRESUMEN
Despite recent advances in understanding microbial diversity in skin homeostasis, the relevance of microbial dysbiosis in inflammatory disease is poorly understood. Here we perform a comparative analysis of skin microbial communities coupled to global patterns of cutaneous gene expression in patients with atopic dermatitis or psoriasis. The skin microbiota is analysed by 16S amplicon or whole genome sequencing and the skin transcriptome by microarrays, followed by integration of the data layers. We find that atopic dermatitis and psoriasis can be classified by distinct microbes, which differ from healthy volunteers microbiome composition. Atopic dermatitis is dominated by a single microbe (Staphylococcus aureus), and associated with a disease relevant host transcriptomic signature enriched for skin barrier function, tryptophan metabolism and immune activation. In contrast, psoriasis is characterized by co-occurring communities of microbes with weak associations with disease related gene expression. Our work provides a basis for biomarker discovery and targeted therapies in skin dysbiosis.
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Dermatitis Atópica/genética , Interacciones Microbiota-Huesped/genética , Microbiota/genética , Psoriasis/genética , Piel/metabolismo , Piel/microbiología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Casos y Controles , Dermatitis Atópica/microbiología , Disbiosis/genética , Femenino , Expresión Génica , Perfilación de la Expresión Génica , Humanos , Masculino , Persona de Mediana Edad , Psoriasis/microbiología , ARN Ribosómico 16S , Adulto JovenRESUMEN
Massive amounts of metagenomics data are currently being produced, and in all such projects a sizeable fraction of the resulting data shows no or little homology to known sequences. It is likely that this fraction contains novel viruses, but identification is challenging since they frequently lack homology to known viruses. To overcome this problem, we developed a strategy to detect ORFan protein families in shotgun metagenomics data, using similarity-based clustering and a set of filters to extract bona fide protein families. We applied this method to 17 virus-enriched libraries originating from human nasopharyngeal aspirates, serum, feces, and cerebrospinal fluid samples. This resulted in 32 predicted putative novel gene families. Some families showed detectable homology to sequences in metagenomics datasets and protein databases after reannotation. Notably, one predicted family matches an ORF from the highly variable Torque Teno virus (TTV). Furthermore, follow-up from a predicted ORFan resulted in the complete reconstruction of a novel circular genome. Its organisation suggests that it most likely corresponds to a novel bacteriophage in the microviridae family, hence it was named bacteriophage HFM.
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Genoma Viral , Metagenoma , Metagenómica , Proteínas Virales/genética , Secuencia de Bases , Análisis por Conglomerados , Biología Computacional/métodos , Humanos , Cadenas de Markov , Metagenómica/métodos , Anotación de Secuencia Molecular , Sistemas de Lectura AbiertaRESUMEN
Psoriasis is driven by focal disruptions of the immune-homeostasis in human skin. Local relapse following cessation of therapy is common and unpredictable, which complicates clinical management of psoriasis. We have previously shown that pathogenic resident T cells accumulate in active and resolved psoriasis, but whether these cells drive psoriasiform tissue reactions is less clear. Here, we activated T cells within skin explants using the pan-T cell activating antibody OKT-3. To explore if T cells induced different tissue response patterns in healthy and psoriasis afflicted skin, transcriptomic analyses were performed with RNA-sequencing and Nanostring. Core tissue responses dominated by IFN-induced pathways were triggered regardless of the inflammatory status of the skin. In contrast, pathways induced by IL-17A, including Defensin beta 2 and keratinocyte differentiation markers, were activated in psoriasis samples. An integrated analysis of IL-17A and IFN-related responses revealed that IL-17 dominated tissue response correlated with early relapse following UVB treatment. Stratification of tissue responses to T cell activation in resolved lesions could potentially offer individualized prediction of disease relapse during long-term immunomodulatory treatment.