Pediatric Emergency Department, Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangzhou, China.
Pediatric Neurology Department, Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangzhou, Guangdong, China.
Virus Res. 2022 Sep;318:198843. doi: 10.1016/j.virusres.2022.198843. Epub 2022 Jun 2.
The effect of structural viral protein 1 (VP1) on neurological damage caused by enterovirus 71 (EV71) infection is unclear. This study aimed to explore the transcriptome changes in EV infected patients and the role of VP1 on the cell secretion pathway of neuron cells.
In our cohort, EV infected patients were enrolled, and RNA-seq analysis was used to evaluate the distinct transcript patterns of cerebrospinal fluid (CSF). The EV71 VP1-overexpressing vector (pEGFP-c3-VP1) was generated and transfected into neuron cells. The relationship between Glutamate Rich 3 (ERICH3) and methyltransferase Zinc Finger CCCH-Type Containing 13 (ZC3H13) and their effect on the serotonin (5-HT) release of neuron cells were explored using small interfering RNA. The expression of ERICH3 and ZC3H13 and concentration of 5-HT were determined using real-time PCR, Western blot, and ELISA, respectively.
The expression of ERICH3 and ZC3H13 were significantly upregulated in EV infected patients with neurological symptoms compared to those without (P < 0.05). The ERICH3 gene had many N6-methyladenosine (m6A) binding sites that can be regulated by m6A modification. Further, the expression of ERICH3 and ZC3H13 were elevated significantly in EV71-VP1 overexpressing neuron cells (P < 0.05). Moreover, ERICH3 or ZC3H13 deficiency could significantly downregulate the release of 5-HT in VP1-overexpressing cells (P < 0.05). Nonetheless, ERICH3 expression was significantly suppressed when ZC3H13 was silenced in neuron cells and vice versa (P < 0.05).
EV71-VP1 can promote 5-HT release by upregulating the expression of ERICH3 and ZC3H13. 5-HT might be a novel therapeutic target for EV71 infection-induced fatal neuronal damage.
肠道病毒 71 型（EV71）感染引起的神经损伤的结构病毒蛋白 1（VP1）的作用尚不清楚。本研究旨在探讨 EV 感染患者的转录组变化以及 VP1 对神经元细胞分泌途径的影响。
本研究纳入 EV 感染患者，采用 RNA 测序分析评估脑脊液（CSF）中独特的转录模式。生成 EV71 VP1 过表达载体（pEGFP-c3-VP1）并转染至神经元细胞。使用小干扰 RNA 探索谷氨酸丰富 3（ERICH3）和含锌指 CCCH 型甲基转移酶 13（ZC3H13）之间的关系及其对神经元细胞 5-羟色胺（5-HT）释放的影响。使用实时 PCR、Western blot 和 ELISA 分别测定 ERICH3 和 ZC3H13 的表达和 5-HT 的浓度。
与无神经症状的 EV 感染患者相比，有神经症状的 EV 感染患者 ERICH3 和 ZC3H13 的表达显著上调（P<0.05）。ERICH3 基因具有许多可通过 m6A 修饰调节的 N6-甲基腺苷（m6A）结合位点。此外，EV71-VP1 过表达神经元细胞中 ERICH3 和 ZC3H13 的表达明显升高（P<0.05）。此外，在 VP1 过表达细胞中，沉默 ERICH3 或 ZC3H13 可显著下调 5-HT 的释放（P<0.05）。然而，在神经元细胞中沉默 ZC3H13 时 ERICH3 的表达明显受到抑制，反之亦然（P<0.05）。
EV71-VP1 可通过上调 ERICH3 和 ZC3H13 的表达促进 5-HT 的释放。5-HT 可能是 EV71 感染引起致命性神经元损伤的新的治疗靶点。
