Comparing transcriptional profiles of type 2 innate lymphoid cells from WT and Rag1 knockout mice
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ABSTRACT: Type 2 inflammation contributes to the pathology of skin diseases such as atopic dermatitis and urticaria. Type 2 innate lymphoid cells are key mediators of skin inflammation by releasing type 2 cytokines in response to certain environmental stimuli. We recently found that Rag1 knockout mice exhibit more pronounced skin inflammation in a mouse model of atopic-dermatitis-like disease, despite lacking adaptive immune cells like T helper 2 cells, implicating a critical role of type 2 innate lymphoid cells in this condition. The goal of this study was to characterize transcriptional differences between WT and Rag KO type 2 innate lymphoid cells to determine if Rag knockout leads to cell-intrinsic alterations in innate lymphoid cell function.
Project description:Type 2 inflammation contributes to the pathology of skin diseases such as atopic dermatitis (AD) and urticaria. We previously found that AD-like inflammation induced by calcipotriol in mice is critically mediated by group 2 innate lymphoid cells (ILC2s), while T cells and B cells are dispensable given development of robust AD-like disease in RAG knockout mice. Indeed, we found that in RAG knockout mice AD-like inflammation was worse, with a proportion of ILC2s displaying increased markers of activation. The goal of this study was to characterize transcriptional and epigenomic changes in ILC populations with a history, or not, of expressing RAG1 in the setting of AD-like skin inflammation.
Project description:Atopic dermatitis and psoriasis are driven by alternate type 2 and type 17 immune responses, but some proteins might be critical to both diseases. We show that a deficiency of the TNF superfamily molecule TWEAK (TNFSF12) in mice results in defective maintenance of atopic dermatitis-specific Th2 and psoriasis-specific Th17 cells in the skin, and impaired expression of disease-characteristic chemokines and cytokines, such as CCL17 and TSLP in atopic dermatitis, and CCL20 and IL-19 in psoriasis. The TWEAK receptor, Fn14, is upregulated in keratinocytes and dermal fibroblasts, and TWEAK induces these cytokines and chemokines alone and in synergy with the signature T helper cytokines of either disease, IL-13 and IL-17. Furthermore, subcutaneous injection of recombinant TWEAK into naïve mice induces cutaneous inflammation with histological and molecular signs of both diseases. TWEAK is therefore a critical contributor to skin inflammation and a possible therapeutic target in atopic dermatitis and psoriasis.
Project description:Hedgehog (Hh) proteins are morphogens which regulate embryonic development and adult tissue homeostasis, with distinct outcomes dependent on the strength and duration of their signals. We show that the Hh signalling pathway modulates the induction and pathology of mouse atopic dermatitis. Sonic hedgehog (Shh) and Hh pathway target genes were upregulated on induction of atopic dermatitis, and the Hh pathway was activated in skin T cells, showing that they respond in vivo to Hh signals secreted from the skin. Shh upregulation reduced skin inflammation in mice, whereas pharmacological Smoothened-inhibition reduced Shh upregulation and exacerbated skin pathology. Hh-signalling to T cells prevented skin inflammation on induction of dermatitis, while inhibition of Hh-mediated transcription in T cells substantially exacerbated the disease. RNA-sequencing analysis of skin CD4+ T cells from mice with chronic atopic dermatitis revealed decreased expression of immune regulatory genes in mice with conditional inhibition of Hh-mediated transcription in T cells, and increased expression of inflammatory and chemokine genes. In contrast, constitutive Hh mediated transcription in T cells led to increased expression of immune regulatory genes in skin CD4+ T cells from mice with chronic atopic dermatitis and protected against inflammation. Hh-mediated transcription in T cells resulted in increased regulatory T (Treg) cells in the periphery and skin of dermatitis-induced mice, and increased TGF-β expression, supporting their immunoregulatory phenotype, whereas, inhibition of T cell specific Hh-mediated transcription, resulted in impaired Treg function, which permitted progression of skin inflammation.
Project description:The aim of this study was to investigate cutaneous cellular and molecular events in the photodermatoses (including solar urticaria and photoaggravated atopic dermatitis) following solar simulated ultraviolet radiation (SSR) exposure, and this dataset comprised the healthy controls (HC). Cutaneous biopsies were taken from unexposed skin and from skin exposed to a single low (physiological) dose of SSR, at 30 minutes, 3 hours, 24 hours and 72 hours post-exposure, in n=6 HC. Biopsies were assessed by immunohistochemistry (n=6 HC) and RNA-sequencing analysis (n=4 HC).
Project description:Type 2 inflammation contributes to the pathology of pruritic skin diseases such as atopic dermatitis (AD). Chronic pruritus of unknown origin (CPUO) is a skin condition of unknown etiology that typically has minimal inflammation. Yet, some patients with CPUO display signs of mildly increased type 2 inflammation. This study aims to compare skin transcriptional profiles in CPUO to healthy skin and to skin from AD, a classical type 2 inflammatory condition.
Project description:Periostin is a matricellular protein known to be alternatively spliced to produce isoforms with a molecular weight of 78-91 kDa. In the extracellular matrix, periostin attach to cell surfaces and induce signaling via integrin-binding and participates in fibrillogenesis to organize collagen in the extracellular space. In the atopic diseases atopic dermatitis and asthma, periostin is known to participate in driving the disease-causing type 2 inflammation. The periostin isoforms expressed in these diseases and the implication of the alternative splicing events are unknown. Here we present two universal assays to map the expression of periostin isoforms on both the transcriptional (RT-qPCR) and translational (PRM-based mass spectrometry) level. We use these assays to study the splice profile of periostin in atopic dermatitis lesions from patients in active treatment vs. normal skin and in in vitro models of atopic dermatitis and asthma. All isoforms expect isoform 3 show decreased expression at the transcriptional level in AD lesions from patients treated with corticosteroids compared to normal skin. The isoforms display an elevated amount at the translational level indicating a delayed response in periostin level during treatment. Expression of the isoforms were upregulated in the in vitro models of atopic dermatitis and asthma at both the transcriptional and translational level with isoform 3 and 5 displaying the highest level of overexpression. Interestingly, the often overlooked isoform 9 and 10 behaved opposite to the other isoforms as they were equally or even less abundant in the disease models compared to the control, and they were identified in the normal skin samples but not in atopic dermatitis lesions. With the assays and findings presented in the publication connected to this dataset we can take further steps in mapping and understanding the role of periostin isoforms.
Project description:Atopic dermatitis, which is a common inflammatory skin disease characterized by persistent epidermal barrier dysfunction, is a systemic health burden reducing overall quality of life of the person. Recently, we showed that the nonstructural extracellular matrix molecule Thrombospondin-4 (THBS4) was upregulated in psoriatic skin lesions by more than 2-fold. In addition, THBS4 contributed to both skin regeneration and wound healing in vitro and in vivo. In the present work we found that THBS4 is also abundantly expressed in atopic dermatitis (AD) patient skin biopsies. By using a proteotransciptomic approach we show that stimulation of primary keratinocytes with THBS4 activates multiple factors, including inflammation, migration, proliferation, keratinocyte differentiation, by which THBS4 could participate in AD progression and contribute to the wound healing process.
Project description:Atopic dermatitis (AD) is the chronic inflammatory skin disease accompanied with severe pruritus. To explore the roles of EGR1 in atopic dermatitis and the relationship between EGR1 and pruritus-scratching behavior, we used a atopic dermatitis-like mouse model driven by house dust mite (HDM) treatment in wild type and EGR1 KO mice, followed with RNA-sequencing analysis.
Project description:Atopic dermatitis (AD) is a common pruritic dermatitis with macroscopically nonlesional skin that is often abnormal. Therefore, we used high-density oligonucleotide arrays to identify cutaneous gene transcription changes associated with early AD inflammation as potential disease control targets. Skin biopsy specimens analyzed included normal skin from five healthy nonatopic adults and both minimally lesional skin and nearby or contralateral nonlesional skin from six adult AD patients. Keywords: disease state analysis We used high-density oligonucleotide Affymetrix Human U133A GeneChip arrays to identify cutaneous gene transcription changes associated with early AD inflammation as potential disease control targets. Skin biopsy specimens analyzed included normal skin from five healthy nonatopic adults and both minimally lesional skin and nearby or contralateral nonlesional skin from six adult AD patients.
Project description:Purpose: Transcriptomic analysis of ADAM17 cKO mice, a mouse model of atopic dermatitis. Methods: Skin was harvested from 10-week-old wild-type and ADAM17 cKO mice and were subjected to next generation sequencing as described. Results: Of 13,573 genes analyzed by GSA, 2514 differentially expressed genes (DEGs) were detected in ADAM17 cKO skin using a standard definition including both statistically significant change (P-value < 0.01; FDR < 0.05) and effect size (|fold change| > 2) . These DEGs included genes important in lipid metabolism and keratinization, consistent with the understanding of barrier dysfunction in atopic inflammation, as well as significant immunologic pathways in adaptive and innate immunity previously identified in the human AD transcriptome Conclusions: Our study reveals that the transcriptome of ADAM17 cKO mice capture a part of upregulated genes previously observed in human AD skin.