Gpr137b is an orphan G-protein-coupled receptor associated with M2 macrophage polarization
Gpr137b is an orphan G-protein-coupled receptor associated with M2 macrophage polarization
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DOI：
10.1016/j.bbrc.2018.12.140
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发表时间：
2019-02-12
影响因子：
3.1
通讯作者：
Ishibashi, Osamu
中科院分区：
文献类型：
--
作者：
Islam, Zohirul;Inui, Takashi;Ishibashi, Osamu
根据其微环境，大型真菌主要分为两个亚型，M1和M2，它们表现出不同的表型。尽管最近的研究表明G蛋白偶联受体（GPCR）与M1/M2巨噬细胞极化相关，但关于GPCR介导的巨噬细胞极化的可用信息仍然有限。在本研究中，我们确定了Gpr 137 b作为孤儿GPCR在小鼠巨噬细胞系RAW 264中大量表达，并阐明了其在M2巨噬细胞极化中的作用。我们使用CRISPR/Cas9基因组编辑系统产生RAW 264细胞的Gpr 137 b敲除（Gpr 137 b-KO）克隆。分离出两个独立的Gpr 137 b-KO克隆，其被证明在含有Gpr 137 b的ATG起始密码子的区域具有移码188个核苷酸缺失。因此，qRT-PCR分析显示缺失的区域不转录。然后，我们用白细胞介素-4（IL-4）处理Gpr 137 b-KO和野生型RAW 264细胞以诱导M2巨噬细胞极化。微阵列分析显示，在Gpr 137 b-KO细胞中，IL-4诱导的代表性M2巨噬细胞标志物的基因表达显著降低，这通过qRT-PCR分析得到验证。相比之下，脂多糖诱导的M1巨噬细胞标志物基因表达不受Gprl 37 b-KO的影响。总的来说，目前的研究表明，Gpr 137 b是M2巨噬细胞极化的可能调节剂。(C)2018爱思唯尔公司All rights reserved.
Macrophages are classified mainly into two subtypes, M1 and M2, which exhibit distinct phenotypes, based on their microenvironment. Although recent studies have suggested that G-protein-coupled receptors (GPCRs) are associated with M1/M2 macrophage polarization, available information on GPCR-mediated macrophage polarization is still limited. In the present study, we identified Gpr137b as an orphan GPCR abundantly expressed in RAW264, a mouse macrophage cell line, and illuminated its role in M2 macrophage polarization. We generated Gpr137b-knockout (Gpr137b-KO) clones of RAW264 cells using the CRISPR/Cas9 genome editing system. Two independent Gpr137b-KO clones were isolated, which were demonstrated to have frameshifting 188-nucleotide deletions at a region containing the ATG start codon of Gpr137b. Consistently, qRT-PCR analysis revealed that the deleted region is not transcribed. We then treated the Gpr137b-KO and wildtype RAW264 cells with interleukin-4 (IL-4) to induce M2 macrophage polarization. Microarray analysis revealed that the IL-4-induced gene expression of representative M2 macrophage markers was significantly reduced in the Gpr137b-KO cells, and this was validated by qRT-PCR analysis. By contrast, M1 macrophage marker gene expression induced by lipopolysaccharide was unaffected by Gprl37b-KO. Collectively, the current study shows that Gpr137b is a possible regulator of M2 macrophage polarization. (C) 2018 Elsevier Inc. All rights reserved.