IgG from systemic sclerosis patients induce a profibrosing and serotype-dependent phenotype in normal dermal fibroblast: a multi-omics study
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE178299 | GEO | 2022/06/07
REPOSITORIES: GEO
Similar Datasets
Project description:Autoantibodies (Aab) are frequent in systemic sclerosis (SSc). While recognized as potent biomarkers, their pathogenic role is much debated. This study explored the effect of purified IgG from SSc patients on the phenotype and function of healthy dermal fibroblast (FB) using an innovative multi-omics approach.
2022-06-29 | PXD025885 | Pride
Project description:Objectives: Eph/Ephrin cell-cell signaling is emerging as a key player in tissue fibrogenesis. The aim of this study was to test the hypothesis that the receptor tyrosine kinase EphB2 mediates dermal fibrosis in systemic sclerosis (SSc). Methods: We assessed normal and SSc human skin biopsies for EphB2 expression. The in vivo role of EphB2 in skin fibrosis was investigated by subjecting EphB2-knockout mice to both bleomycin-induced and tight skin (Tsk1/+) genetic mouse models. EphB2 kinase-dead and overactive point mutant mice were used to evaluate the role of EphB2 forward signaling in bleomycin-induced dermal fibrosis. In vitro studies were performed on dermal fibroblasts from SSc patients and healthy controls, which was followed by in vivo analysis of fibroblast-specific EphB2 deficient mice. Results: Expression of EphB2 is upregulated in SSc skin tissue and explanted SSc dermal fibroblasts compared to healthy controls. EphB2 expression is elevated in two animal models of dermal fibrosis. In mice, EphB2 drives dermal fibrosis in both the bleomycin and the Tsk1/+ models of skin fibrosis. EphB2 forward signaling is a critical mediator of dermal fibrosis. Transforming growth factor-β (TGFβ) cytokines upregulate EphB2 in dermal fibroblasts via non-canonical TGFβ/SMAD signaling and silencing EphB2 in dermal fibroblasts is sufficient to dampen TGFβ-induced fibroblast-to-myofibroblast differentiation. Moreover, mice with fibroblast-specific deletion of EphB2 showed impaired fibroblast-to-myofibroblast differentiation and reduced skin fibrosis upon bleomycin challenge. Conclusion: Our data implicate EphB2 overexpression and kinase-mediated forward signaling in the development of dermal fibrosis in SSc. EphB2 thus represents a potential new therapeutic target for SSc.
2024-04-10 | GSE253089 | GEO
Project description:Systemic sclerosis (SSc) is a heterogeneous connective tissue disease characterized by autoimmunity, vasculopathy and excessive fibrosis. Antinuclear antibodies (ANA) are strong diagnostic and prognosis biomarkers but their pathogenicity in fibrosis and vasculopathy remains unclear. Functional antibodies (FA) have been described and might be implicated in the vasculopathy of SSc, in which endothelial cells (EC) are key players. We aimed to explore the effect of purified IgG from SSc patients on omics signature of EC and examine the influence of ANA serotypes or FA.
2025-03-24 | PXD057210 | Pride
Project description:Systemic sclerosis (SSc) is a heterogeneous connective tissue disease characterized by autoimmunity, vasculopathy and excessive fibrosis (1). Antinuclear antibodies (ANA) are strong diagnostic and prognosis biomarkers but their pathogenicity in fibrosis and vasculopathy remains unclear. Functional antibodies (FA) such as angiotensin II type 1 receptor [AT1R] and endothelin-1 type A receptor [ETAR] have been described in SSc and might be implicated in vasculopathy, in which endothelial cells (EC) are key players.We aimed to explore the effect of purified IgG from SSc patients on the proteome of EC and examine the influence of ANA serotypes or FA. The detailed methodology is available in the Supplementary Material section.
2025-03-24 | PXD049364 | Pride
Project description:IgG from systemic sclerosis patients induce a profibrosing and serotype-dependent phenotype in normal dermal fibroblast: a multi-omics study
| PRJNA738391 | ENA
Project description:Background. The etiology and reasons underlying the ethnic disparities in systemic sclerosis (SSc) remain unknown. African-Americans (AA) are disproportionally affected by SSc, yet dramatically underrepresented in research. The role of DNA methylation in disease risk remains unclear. This analysis was conducted to comprehensive identify differentially methylated loci associated with SSc in AA. Methods. Genomic DNA was isolated from cultured dermal fibroblasts isolated from 15 AA SSc cases and 15 AA controls. All patients met the 2013 ACR/EULAR classification criteria for SSc, most (93%) presenting with diffuse cutaneous SSc. DNA methylation patterns were profiled through reduced representation bisulfite sequencing (RRBS). Alignment and methylation calling were performed using Bismarck v0.16.3 and the GRCh37/hg19 reference genome. Data was filtered, normalized, and analyzed with RnBeads v1.6.1. Differential methylation analysis was conducted on CpG, promoter, gene and system level. Results. We generated DNA methylation data for over 5 million CpGs in each sample and between 10x to 40x coverage in CpGs sites. Using the Combined Score approach implemented in RnBeads, a total of 9 CpG islands, 17 genes and 11 promoters showed significant differential methylation levels between cases and controls at the gene-level. The top differentially methylated genes constitute mostly non-coding RNA genes (42%), followed by pseudogenes (27%), then protein coding genes (19%). Enrichment analysis revealed that both hypo- and hypermethylated genes and their promoter regions were enriched for cell differentiation and immune-related gene ontology terms (hypermethylated regions: mesenchymal cell transition and differentiation; hypomethylated regions: type I IFN signaling). Conclusion. The observed DNA methylation differences between cases and controls support a modest role for DNA methylation differences in mediating susceptibility to SSc in AA. Instead of protein-coding, most differentially methylated genes comprise non-coding RNA genes, supporting a larger contribution of dysregulated regulatory elements to disease. While previous DNA methylation profiling analyses in European-Americans reported an enrichment of extracellular matrix and focal adhesion genes, our data supports a potentially stronger immune-driven etiology in AA.
2021-01-19 | GSE150592 | GEO
Project description:ObjectivesPrior work demonstrates that co-cultured macrophages and fibroblasts from patients with SSc engage in reciprocal activation. However, the mechanism by which these cell types communicate and contribute to fibrosis and inflammation in SSc is unknown.MethodsFibroblasts were isolated from skin biopsies obtained from 7 SSc patients or 6 healthy age and gender-matched control subjects following written informed consent. Human donor-derived macrophages were cultured with exosomes isolated from control or SSc fibroblasts for an additional 48 h. Macrophages were immunophenotyped using flow cytometry, qRT-PCR and multiplex. For mutual activation studies, exosome-activated macrophages were co-cultured with SSc or healthy fibroblasts using Transwells.ResultsMacrophages activated with dermal fibroblast-derived exosomes from SSc patients upregulated surface expression of CD163, CD206, MHC Class II and CD16 and secreted increased levels of IL-6, IL-10, IL-12p40 and TNF compared with macrophages incubated with healthy control fibroblasts (n = 7, P < 0.05). Exosome-stimulated macrophages and SSc fibroblasts engaged in reciprocal activation, as production of collagen and fibronectin was significantly increased in SSc fibroblasts receiving signals from SSc exosome-stimulated macrophages (n = 7, P < 0.05).ConclusionIn this work, we demonstrate for the first time that human SSc dermal fibroblasts mediate macrophage activation through exosomes. Our findings suggest that macrophages and fibroblasts engage in cross-talk in SSc skin, resulting in mutual activation, inflammation, and extracellular matrix (ECM) deposition. Collectively, these studies implicate macrophages and fibroblasts as cooperative mediators of fibrosis in SSc and suggest therapeutic targeting of both cell types may provide maximal benefit in ameliorating disease in SSc patients.
| S-EPMC9910573 | biostudies-literature
Project description:Aberrant deposition of extracellular matrix (ECM) resulting in dermal fibrosis is a hallmark of systemic sclerosis (SSc). Evidence suggests that dysfunctional SSc keratinocytes may contribute to fibrosis by altering dermal homeostasis. Whether interleukin-25 (IL-25), an IL-17 family member involved in epithelial/mesenchymal/immune cell interplay takes part in skin fibrosis is unknown. Here we address the role of IL-25 in SSc skin fibrosis. Compared to healthy donor (HD), in SSc and scleroderma-like disorders the epidermis expressed significantly lower levels of IL-25. In epidermal equivalents, IL-25 regulated several molecular pathways related to wound healing and ECM remodeling. Compared to control conditioned medium (CM), the CM from IL-25-primed keratinocytes enhanced the production by fibroblasts of matrix metalloproteinase-1 (MMP-1), IL-6, IL-8 (p< 0.05), but not of type-I collagen (Col-I ) nor fibronectin. However, IL- 25 significantly reduced the production of Col-I when applied directly to fibroblasts. The activation of keratinocytes by IL-25 was receptor-dependent and evident after a very short incubation time (10 min), largely mediated by IL-1, suggesting enhanced and specific release of preformed mediators. These results show that IL-25 participates to skin homeostasis and its decreased expression in SSc may contribute to skin fibrosis by favoring ECM deposition over degradation.
2022-03-31 | GSE168312 | GEO
Project description:Systemic sclerosis (SSc) is characterized by systemic fibrosis, vascular dysfunction, and the presence of antinuclear autoantibodies. In up to 10% of persons with SSc, the target antigens of the antinuclear autoantibodies are unknown. Using IP-MS, we here describe another new target antigen in SSc. We examined a database of IP-MS identifications of 106 persons with SSc from two Belgian academic hospitals [Ghent University Hospital (n=59) and University Hospitals Leuven (n=47)] without autoantibody specificity as tested by commercial assays.TATA-binding protein (TBP)-associated factors (TAFs) were precipitated by sera of four persons (two from Ghent, two from Leuven). Together with TBP, TAFs form the three-lobed transcription factor IID complex (TFIID) (figure 1B). The TFIID complex recognizes core promoter sequences on DNA. The binding of TFIID is the first step of the assembly of the preinitiation complex, which is needed to position RNA polymerase II onto the correct DNA sequence to start transcription. To identify additional cases, we tested samples from individuals with SSc with unidentified autoantibodies from 3 extra hospitals [Cliniques Universitaires Saint-Luc (Belgium) (n=45), Chris Hani Baragwanath Hospital (South Africa) (n=28) and Centre Hospitalier Universitaire d’Angers (France) (n=13)] by IP-MS. Two additional cases precipitating TFIID subunits were identified. In order to confirm the specificity of anti-TIFIID antibodies for SSc, IP-MS was performed in healthy individuals (n=34) and in patients with idiopathic inflammatory myopathy (IIM, n=12), Sjögren’s disease (SjD, n=12), systemic lupus erythematosus (SLE, n=36) and anti-topoisomerase-I-, anti-centromere- or anti-RNA polymerase III-positive SSc (n=28). TAF9, TAF9B and TAF10 were often precipitated in these control samples. In conclusion, we describe TFIID as a new target of autoantibodies in SSc, thereby expanding the spectrum of SSc-associated autoantigens related to transcription.
2025-07-01 | PXD064257 | Pride
Project description:Systemic scleroderma (SSc) is an autoimmune disease which results in fibrotic production in the lung. Resultant SSC-pulmonary fibrosis is the main cause of mortality among SSc patients. From high throughput RNAi screening, we uncovered the ubiquitin E3 ligase KLHL42 as a potential pro-fibrotic mediator of TGFb-dependent fibrotic signaling in primary SSc lung fibroblasts. In this analysis, we sought to uncover putative substrates for KLHL42 by comparing SSc lung fibroblasts with control or KLHL42 siRNA prior to TGFb-treatment, lysis, and TUBE precipitation. The resultant pull-down was analyzed with LC-MS/MS.
2020-01-14 | MSV000084800 | MassIVE