The Autoimmune Susceptibility GeneC5orf30Regulates Macrophage-Mediated Resolution of Inflammation

Abstract

Genetic variants in C5orf30 have been associated with development of the autoimmune conditions primary biliary cirrhosis and rheumatoid arthritis. In rheumatoid arthritis, C5orf30 expression is cell-specific, with highest expression found in macrophages and synovial fibroblasts. C5orf30 is highly expressed in inflamed joints and is a negative regulator of tissue damage in a mouse model of inflammatory arthritis. Transcriptomic analysis from ultrasound-guided synovial biopsy of inflamed joints in a well characterized clinical cohort of newly diagnosed, disease-modifying antirheumatic drugs-naive rheumatoid arthritis patients was used to determine the clinical association of C5orf30 expression with disease activity. A combined molecular and computational biology approach was used to elucidate C5orf30 function in macrophages both in vitro and in vivo. Synovial expression of C5orf30 is inversely correlated with both clinical measures of rheumatoid arthritis disease activity and with synovial TNF mRNA expression. C5orf30 plays a role in regulating macrophage phenotype and is differentially turned over in inflammatory and anti-inflammatory macrophages. Inhibition of C5orf30 reduces wound healing/repair-associated functions of macrophages, reduces signaling required for resolution of inflammation, and decreases secretion of anti-inflammatory mediators. In an animal model of wound healing (zebrafish), C5orf30 inhibition increases the recruitment of macrophages to the wound site. Finally, we demonstrate that C5orf30 skews macrophage immunometabolism, demonstrating a mechanism for C5orf30-mediated immune regulation.

Genomic Diversity, Population Structure, and Signature of Selection in Five Chinese Native Sheep Breeds Adapted to Extreme Environments

“…Multiples candidate genes were also found to participate in host defense mechanism, resistance diseases, and inflammatory responses, with two most significant genes worth to mention as examples. C5orf30 gene was reported in association with the development of autoimmune disorders in primary biliary cirrhosis as well as the susceptibility and severity of rheumatoid arthritis in humans [55,56]. Another interesting gene is ITCH, which encodes a member of the Nedd4 family of HECT domain E3 ubiquitin ligases and plays a role in protein trafficking and immune response, and in several signaling pathways that regulate cellular growth and proliferation in multiple processes [57].…”

Association of the Rheumatoid Arthritis Severity Variant rs26232 with the Invasive Activity of Synovial Fibroblasts

“…Total mRNA was extracted from cultured RASF and reverse-transcribed. Reverse transcription of 100 ng of total mRNA was carried out using a high-capacity cDNA reverse transcription kit (Applied Biosystems, Waltham, MA, USA), as previously described [6]. Real-time PCR was carried out on a Roche Lightcycler (Basel, Switzerland) under standard cycling conditions.…”

“…Inhibition of C5orf30 in RASFs results in increased invasive and migratory activities in vitro, and in the collagen-induced arthritis model results in more severe inflammation and joint damage [5]. In macrophages, proinflammatory signals such as tumour necrosis factor (TNF) and lipopolysaccharide (LPS) reduce C5orf30 expression via a c-Jun N-terminal kinase mediated mechanism; furthermore, C5orf30 enhances the resolution of inflammation and wound repair functions mediated by M2 macrophages [6].…”

The transcription factor CUX1 negatively regulates invasion in castrate resistant prostate cancer

“…Whole cell protein lysates were extracted as previously described [38]. Nuclear and cytosolic fractions were extracted from fresh cell pellets washed in PBS, lysed in ice-cold cytosolic buffer (10 mM HEPES pH8.0, 1.5 mM MgCl 2 , 10 mM KCL, 200 mM sucrose, 0.25%NP40, 5 mM DTT, 1× protease inhibitor cocktail (Sigma-Aldrich, MO, USA), 1× phosphatase inhibitor cocktail 1 (Sigma-Aldrich, MO, USA)), harvested to tubes and incubated at 4° C with shaking for 30 minutes followed by centrifugation at 14,000 rpm × 10 mins at 4° C. Supernatant containing cytosolic protein fraction were kept on ice and pellets washed with ice-cold PBS.…”