Assadi Ghazaleh, Vesterlund Liselotte, Bonfiglio Ferdinando, Mazzurana Luca, Cordeddu Lina, Schepis Danika, Mjösberg Jenny, Ruhrmann Sabrina, Fabbri Alessia, Vukojevic Vladana, Percipalle Piergiorgio, Salomons Florian A, Laurencikiene Jurga, Törkvist Leif, Halfvarson Jonas, D'Amato Mauro
Department of Biosciences and Nutrition, Karolinska Institutet, Stockholm, Sweden.
Center for Infectious Medicine, Department of Medicine Huddinge, Karolinska Institutet, Stockholm, Sweden.
PLoS One. 2016 Dec 13;11(12):e0168276. doi: 10.1371/journal.pone.0168276. eCollection 2016.
Genetic variation in the Laccase (multicopper oxidoreductase) domain-containing 1 (LACC1) gene has been shown to affect the risk of Crohn's disease, leprosy and, more recently, ulcerative colitis and juvenile idiopathic arthritis. LACC1 function appears to promote fatty-acid oxidation, with concomitant inflammasome activation, reactive oxygen species production, and anti-bacterial responses in macrophages. We sought to contribute to elucidating LACC1 biological function by extensive characterization of its expression in human tissues and cells, and through preliminary analyses of the regulatory mechanisms driving such expression.
We implemented Western blot, quantitative real-time PCR, immunofluorescence microscopy, and flow cytometry analyses to investigate fatty acid metabolism-immune nexus (FAMIN; the LACC1 encoded protein) expression in subcellular compartments, cell lines and relevant human tissues. Gene-set enrichment analyses were performed to initially investigate modulatory mechanisms of LACC1 expression. A small-interference RNA knockdown in vitro model system was used to study the effect of FAMIN depletion on peroxisome function.
FAMIN expression was detected in macrophage-differentiated THP-1 cells and several human tissues, being highest in neutrophils, monocytes/macrophages, myeloid and plasmacytoid dendritic cells among peripheral blood cells. Subcellular co-localization was exclusively confined to peroxisomes, with some additional positivity for organelle endomembrane structures. LACC1 co-expression signatures were enriched for genes involved in peroxisome proliferator-activated receptors (PPAR) signaling pathways, and PPAR ligands downregulated FAMIN expression in in vitro model systems.
FAMIN is a peroxisome-associated protein with primary role(s) in macrophages and other immune cells, where its metabolic functions may be modulated by PPAR signaling events. However, the precise molecular mechanisms through which FAMIN exerts its biological effects in immune cells remain to be elucidated.
含漆酶(多铜氧化还原酶)结构域1(LACC1)基因的遗传变异已被证明会影响克罗恩病、麻风病的发病风险,最近还发现其与溃疡性结肠炎和青少年特发性关节炎有关。LACC1的功能似乎是促进脂肪酸氧化,同时激活炎性小体、产生活性氧并引发巨噬细胞的抗菌反应。我们试图通过广泛表征其在人体组织和细胞中的表达,并对驱动这种表达的调控机制进行初步分析,来阐明LACC1的生物学功能。
我们采用蛋白质免疫印迹、定量实时聚合酶链反应、免疫荧光显微镜和流式细胞术分析,研究脂肪酸代谢-免疫关联蛋白(FAMIN;LACC1编码的蛋白质)在亚细胞区室、细胞系和相关人体组织中的表达。进行基因集富集分析以初步研究LACC1表达的调节机制。使用体外小干扰RNA敲低模型系统研究FAMIN缺失对过氧化物酶体功能的影响。
在巨噬细胞分化的THP-1细胞和几种人体组织中检测到FAMIN表达,在外周血细胞中,中性粒细胞、单核细胞/巨噬细胞、髓样和浆细胞样树突状细胞中的表达最高。亚细胞共定位仅局限于过氧化物酶体,细胞器内膜结构也有一些额外的阳性信号。LACC1共表达特征富集了参与过氧化物酶体增殖物激活受体(PPAR)信号通路的基因,并且在体外模型系统中PPAR配体下调了FAMIN的表达。
FAMIN是一种与过氧化物酶体相关的蛋白质,在巨噬细胞和其他免疫细胞中起主要作用,其代谢功能可能受PPAR信号事件调节。然而,FAMIN在免疫细胞中发挥生物学效应的确切分子机制仍有待阐明。