Project description:TWEAK, also known as TNFSF12, is a multifunctional cytokine that controls many cellular activities in tissue repair and disease by binding to its receptor Fn14.TWEAK-Fn14 signaling has been shown to induce canonical and noncanonical NFkB signaling, triggering pro-inflammatory responses and leukocyte infiltration.We found that TWEAK-Fn14 signaling was upregulated in IPF while Fn14 was mainly expressed in fibroblasts. Therefore, we added different concentrations of TWEAK to primary mouse lung fibroblasts to activate the TWEAK-Fn14 signaling pathway, and explored the transcriptome changes of TWEAK-Fn14 signaling pathway activation in fibroblasts by RNA-seq technology.
Project description:Fibroblasts acquire a pro-inflammatory phenotype in inflammatory bowel disease (IBD), but the factors driving this process and how fibroblasts contribute to the immune response is incompletely understood. The TNF superfamily factor 12 (TWEAK) has gained interest as a mediator of chronic inflammation. Here, we explore its role as a driver of inflammatory responses in fibroblasts and its contribution to fibroblast-monocyte interaction using human primary colonic fibroblasts, THP1 and peripheral blood mononuclear cells (PBMC). TWEAK induced the expression of cytokines, chemokines and immune receptors in fibroblasts. The TWEAK up-regulated transcriptome shared 29% homology with the previously published transcriptional profile of inflammatory fibroblasts from ulcerative colitis. TWEAK elevated surface expression of activated fibroblasts markers and adhesion molecules (PDPN, ICAM-1 and VCAM-1) and secretion of IL-6, CCL2 and CXCL10. In co-culture, fibroblasts induced monocyte adhesion and promoted a CD14high/ICAM-1 high phenotype in THP1 cells, both of which were enhanced when fibroblasts were pre-stimulated with TWEAK. Medium from TWEAK-treated fibroblast-THP1 co-cultures had elevated levels of CXCL1 and IL-8. PBMCs in co-culture with TWEAK-treated fibroblasts had altered surface expression of CCR2 and CD16, and increased CXCL1 and CXCL10. Our results indicate that TWEAK promotes and inflammatory fibroblast-monocyte crosstalk that may be amenable for therapeutic intervention.
Project description:The expression of TWEAK and Fn14 was increased in early skin lesions of SSc patients. Fn14 expression on human dermal fibroblasts is significantly increased after stimulation with serum from patients with systemic sclerosis. The interplay between TWEAK and Fn14 is pivotal in the fibrotic progression of various autoimmune diseases. These observations implicate the TWEAK/Fn14 signaling pathway may as a critical player in the pathological advancement of systemic sclerosis; however, the precise underlying mechanisms require further clarification. Thus, human dermal fibroblasts were stimulated by serum from systemic sclerosis patients, and with or without the addition of Fn14 antagonists. We then assessed the antifibrotic effects of this antagonist through RNA sequencing, and preliminary exploration of possible molecular mechanisms.
Project description:To investigate the transcriptome of TWEAK or TGF-α in the regulation of human primary keratinocytes We then performed gene expression profiling analysis using data obtained from RNA-seq of 3 replicates at two different time points
Project description:Pulmonary fibrosis (PF) is both an independent disease and a pathologic basis for fibroproliferative lung diseases. Understanding of underlying mechanisms of PF is critical for developing effective therapeutics. Fibroblasts activation and extracellular matrix deposition are critical in pathogenesis of PF. Stimulation of certain factors can induce profibrotic changes in fibroblasts. Evaluation of associated genes in treated fibroblasts is helpfu to analyze the profibrotic changes of this critical effector cell for PF. We used microarrays to detail the global programme of gene expression following TGFβ and CCL1 stimulation of primary lung fibroblasts.
Project description:We used microarrays to profile gene expression changes following growth factor stimulation of primary human fibroblasts. We serum starved (0.1% serum) fibroblasts for 48 hrs and restimulated with 10% serum for 0, 2, 4, 6 and 8 hrs. Total RNA was extracted from 2 independent biological replicas for each time point and hybridized to expression arrays.
Project description:Tissue fibrosis is a common pathway to organ injury and failure. It is characterized by an excessive deposition of extracellular matrix (ECM) in organs. Deciphering the fibrogenic processes is of utmost importance, as there are few effective therapies in fibrotic diseases 1. Systemic sclerosis (SSc) is a prototypical disease where fibroblasts (Fb) are key effector cells as they differentiate into myofibroblasts in response to chronic inflammation under the influence of transforming growth factor beta 1 (TGF-β1) pathway 2–4. In this study, we compared the proteome of primary Fb in different culture and stimulation conditions. Primary dermal normal human Fb were cultured at passage P3, P5 and P7 with and without Fetal Bovine Serum (FBS). At fifth passage, Fb were stimulated or not with different concentrations of recombinant human active TGF-β1 (0.04, 1 and 5 ng/mL) during 24, 48 and 72 hours.