Demethylation of G-Protein-Coupled Receptor 151 Promoter Facilitates the Binding of Kruppel-Like Factor 5 and Enhances Neuropathic Pain after Nerve Injury in Mice
Demethylation of G-Protein-Coupled Receptor 151 Promoter Facilitates the Binding of Kruppel-Like Factor 5 and Enhances Neuropathic Pain after Nerve Injury in Mice
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G 蛋白偶联受体 151 启动子的去甲基化促进 Krüppel 样因子 5 的结合并增强小鼠神经损伤后的神经病理性疼痛
DOI：
10.1523/jneurosci.0702-18.2018
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发表时间：
2018-12-05
影响因子：
5.3
通讯作者：
Gao, Yong-Jing
中科院分区：
文献类型：
--
作者：
Jiang, Bao-Chun;Zhang, Wen-Wen;Gao, Yong-Jing
G蛋白偶联受体被认为是细胞外信号的细胞表面传感器，因此在信号转导中具有关键作用，并且是药物发现的最富有成效的靶点。G蛋白偶联受体151（GPR151）被报道在缰核区特异性表达。本文报道了GRP151在脊神经结扎（SNL）后脊髓中的表达和表观遗传调控，以及GRP151在神经病理性疼痛中的作用。SNL显著增加脊髓神经元中GPR151的表达。GPR151突变或通过shRNA抑制脊髓减轻SNL诱导的机械异常性疼痛和热痛觉过敏。SNL后GPR151基因启动子区的CpG岛发生脱甲基化，DNA甲基转移酶3b（DNMT3b）的表达降低，DNMT3b与GPR151启动子的结合减少。脊髓中DNMT 3b的过表达降低了GPR151的表达并减轻了SNL诱导的神经病理性疼痛。此外，Kruppel样因子5（KLF 5），KLF家族的转录因子，在脊髓神经元中上调，KLF 5与GPR151启动子的结合亲和力增加后SNL。抑制KLF5可降低GPR151的表达，并减弱SNL诱导的疼痛超敏反应。进一步的mRNA微阵列分析显示，GPR151的突变降低了SNL引起的多种疼痛相关基因的表达，尤其是丝裂原活化蛋白激酶（MAPK）信号通路相关基因。这项研究表明，GPR151，增加DNA去甲基化和增强与KLF5的相互作用，有助于通过增加MAPK通路相关的基因表达的神经病理性疼痛的维持。
G-protein-coupled receptors are considered to be cell-surface sensors of extracellular signals, thereby having a crucial role in signal transduction and being the most fruitful targets for drug discovery. G-protein-coupled receptor 151 (GPR151) was reported to be expressed specifically in the habenular area. Here we report the expression and the epigenetic regulation of GRP151 in the spinal cord after spinal nerve ligation (SNL) and the contribution of GPR151 to neuropathic pain in male mice. SNL dramatically increased GPR151 expression in spinal neurons. GPR151 mutation or spinal inhibition by shRNA alleviated SNL-induced mechanical allodynia and heat hyperalgesia. Interestingly, the CpG island in the GPR151 gene promoter region was demethylated, the expression of DNA methyltransferase 3b (DNMT3b) was decreased, and the binding of DNMT3b with GPR151 promoter was reduced after SNL. Overexpression of DNMT3b in the spinal cord decreased GPR151 expression and attenuated SNL-induced neuropathic pain. Furthermore, Kruppel-like factor 5 (KLF5), a transcriptional factor of the KLF family, was upregulated in spinal neurons, and the binding affinity of KLF5 with GPR151 promoter was increased after SNL. Inhibition of KLF5 reduced GPR151 expression and attenuated SNL-induced pain hypersensitivity. Further mRNA microarray analysis revealed that mutation of GPR151 reduced the expression of a variety of pain-related genes in response to SNL, especially mitogen-activated protein kinase (MAPK) signaling pathway-associated genes. This study reveals that GPR151, increased by DNA demethylation and the enhanced interaction with KLF5, contributes to the maintenance of neuropathic pain via increasing MAPK pathway-related gene expression.