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Anti-inflammatory effects of ?-MSH through p-CREB expression in sarcoidosis like granuloma model.

ABSTRACT: Lung inflammation due to sarcoidosis is characterized by a complex cascade of immunopathologic events, including leukocyte recruitment and granuloma formation. ?-melanocyte stimulating hormone (?-MSH) is a melanocortin signaling peptide with anti-inflammatory properties. We aimed to evaluate the effects of ?-MSH in a novel in vitro sarcoidosis model. An in vitro sarcoidosis-like granuloma model was developed by challenging peripheral blood mononuclear cells (PBMCs) derived from patients with confirmed treatment-naïve sarcoidosis with microparticles generated from Mycobacterium abscessus cell walls. Unchallenged PBMCsand developed granulomas were treated daily with 10??M ?-MSH or saline as control. Cytokine concentrations in supernatants of culture and in cell extracts were measured using Illumina multiplex Elisa and western blot, respectively. Gene expression was analyzed using RNA-Seq and RT-PCR. Protein secretion and gene expression of IL-7, IL-7R, IFN-?, MC1R, NF-?B, phosphorylated NF-?B (p-NF-?B), MARCO, and p-CREB were measured with western blot and RNAseq. A significant increase in IL-7, IL-7R, and IFN-? protein expression was found in developed granulomas comparing to microparticle unchallenged PBMCs. IL-7, IL-7R, and IFN-? protein expression was significantly reduced in developed granulomas after exposure to ?-MSH compared with saline treated granulomas. Compared with microparticle unchallenged PBMCs, total NF-?B and p-NF-?B were significantly increased in developed granulomas, while expression of p-CREB was not changed. Treatment with ?-MSH promoted a significantly higher concentration of p-CREB in granulomas. The anti-inflammatory effects of ?-MSH were blocked by specific p-CREB inhibition. ?-MSH has anti-inflammatory properties in this in vitro granuloma model, which is an effect mediated by induction of phosphorylation of CREB.

SUBMITTER: Zhang C

PROVIDER: S-EPMC7190699 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature

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Anti-inflammatory effects of α-MSH through p-CREB expression in sarcoidosis like granuloma model.

Zhang Chongxu C Chery Stephanie S Lazerson Aaron A Altman Norman H NH Jackson Robert R Holt Greg G Campos Michael M Schally Andrew V AV Mirsaeidi Mehdi M

Scientific reports 20200429 1

Lung inflammation due to sarcoidosis is characterized by a complex cascade of immunopathologic events, including leukocyte recruitment and granuloma formation. α-melanocyte stimulating hormone (α-MSH) is a melanocortin signaling peptide with anti-inflammatory properties. We aimed to evaluate the effects of α-MSH in a novel in vitro sarcoidosis model. An in vitro sarcoidosis-like granuloma model was developed by challenging peripheral blood mononuclear cells (PBMCs) derived from patients with con ...[more]

PMID: 32350353

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Project description:The mechanisms underlying abnormal granuloma formation in patients with sarcoidosis are complex and remain poorly understood. A novel in vitro human granuloma model was used to determine the molecular mechanisms of granuloma genesis in patients with sarcoidosis in response to putative disease-causing mycobacterial antigens. Peripheral blood mononuclear cells (PBMCs) from patients with active sarcoidosis and from normal, disease-free control subjects were incubated for 7 days with purified protein derivative-coated polystyrene beads. Molecular responses, as reflected by differential expression of genes, extracellular cytokine patterns, and cell surface receptor expression, were analyzed. Unbiased systems biology approaches were used to identify signaling pathways engaged during granuloma formation. Model findings were compared with human lung and mediastinal lymph node gene expression profiles. Compared with identically treated PBMCs of control subjects (n = 5), purified protein derivative-treated sarcoidosis PBMCs (n = 6) were distinguished by the formation of cellular aggregates resembling granulomas. Ingenuity Pathway Analysis of differential expression gene patterns identified molecular pathways that are primarily regulated by IL-13, which promotes alternatively activated (M2) macrophage polarization. M2 polarization was further demonstrated by immunohistochemistry performed on the in vitro sarcoidosis granuloma-like structures. IL-13-regulated gene pathways were confirmed in human sarcoidosis lung and mediastinal lymph node tissues. The in vitro human sarcoidosis granuloma model provides novel insights into early granuloma formation, particularly IL-13 regulation of molecular networks that regulate M2 macrophage polarization. M2 macrophages are predisposed to aggregation and multinucleated giant cell formation, which are characteristic features of sarcoidosis granulomas. Clinical trial registered with www.clinicaltrials.gov (NCT01857401).

Dataset Information

Anti-inflammatory effects of ?-MSH through p-CREB expression in sarcoidosis like granuloma model.

Publications

Anti-inflammatory effects of α-MSH through p-CREB expression in sarcoidosis like granuloma model.

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