Dataset Information

Culture and stimulation conditions influence protein expression of primary fibroblasts

ABSTRACT: 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.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Fibroblast Of Dermis, Cell Culture

DISEASE(S): Systemic Scleroderma

SUBMITTER: BRAY FABRICE

LAB HEAD: Fabrice Bray

PROVIDER: PXD025374 | Pride | 2021-11-03

REPOSITORIES: Pride

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Passages in culture and stimulation conditions influence protein expression of primary fibroblasts.

Vivier Solange S Chepy Aurélien A Bray Fabrice F Guerrier Thomas T Speca Silvia S Flament Stéphanie S Jendoubi Manel M Balden Maïté M Rolando Christian C Hachulla Eric E Launay David D Dubucquoi Sylvain S Sobanski Vincent V

Proteomics 20211102 3

Fibroblasts (Fb) are key effector cells in systemic sclerosis (SSc). Fb stimulation with transforming growth factor beta 1 (TGF-β1) is considered as a positive control in studies assessing fibrogenesis. The lack of standardization of TGF-β1 stimulation might be responsible for discrepancies in experiments performed in different conditions. Using quantitative proteomics analysis, we evaluated the impact of changes in experimental conditions on proteomic profiles of primary Fb. Principal component ...[more]

PMID: 34665929

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Project description:Background: Increased Galectin 3-binding protein (Lgals3bp) serum levels have been used to assess hepatic fibrosis stages and the severity of hepatocellular carcinoma (HCC). Considering the crucial role of transforming growth factor-β1 (TGF-β1) in the emergence of these diseases, the present study tested the hypothesis that Lgals3bp regulates the TGF-β1 signaling pathway. Methods: The expression levels of Lgals3bp and TGF-β1 were analyzed in patients with non-alcoholic steatohepatitis (NASH) and HCC. Multiple omics techniques, such as RNA-sequencing, transposase-accessible chromatin-sequencing assay, and liquid chromatography-tandem mass spectrometry proteomics, were used to identify the regulatory mechanisms for the Lgals3bp-TGF-β1 axis. Moreover, the effects of altered TGF-β1 signaling in chronic inflammatory conditions were investigated in conditional Lgals3bp-knockin and Lgals3bp-knockout mice. Results: In patients with NASH and HCC, the levels of Lgals3bp and TGF-β1 exhibited positive correlations. Stimulation of Lgals3bp by the inflammatory cytokine interferon α in HCC cells or ectopic overexpression of Lgals3bp in hepatocytes promoted the expression levels of TGFB1. Aggravated fibrosis was observed in the livers of hepatocyte-specific Lgals3bp knock-in mice, with increased TGF-β1 levels. Lgals3bp directly bound to and assembled integrin αV, an integral mediator required for releasing active TGF-β1 from extracellular latent complex with the rearranged F-actin cytoskeleton. The released TGF-β1 activated JunB transcription factor, which in turn promoted the TGF-β1 positive feedback loop. Lgals3bp deletion in the hepatocytes downregulated TGF-β1 signaling and CCl4 induced fibrosis. Finally, Lgals3bp depletion hindered hepatocarcinogenesis by limiting the availability of fibrogenic TGF-β1. Conclusion: Lgals3bp plays a crucial role in hepatic fibrosis and carcinogenesis by controlling the TGF-β1 signaling pathway, making it a promising therapeutic target in TGF-β1-related diseases.

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	004-24-1-1.msf	Msf
	004-24-1-1.raw	Raw
	004-24-1-2.raw	Raw
	004-24-1-3.raw	Raw
	004-48-1-1.msf	Msf