Zhao Yanan, Wang Yuanmei, Liu Liying, Ren Yanru, Liu Long, Wang Jiayi, Li Xianyao
Shandong Provincial Key Laboratory for Livestock Germplasm Innovation & Utilization, College of Animal Science and Technology, Shandong Agricultural University, Tai'an, Shandong, China.
College of Life Sciences, Shandong Agricultural University, Tai'an, Shandong, China.
Front Immunol. 2025 Jul 30;16:1630008. doi: 10.3389/fimmu.2025.1630008. eCollection 2025.
INTRODUCTION: , a commensal food-borne pathogen, poses severe threat to human health and poultry industry. N6-methyladenosine (mA) mRNA modification is associated with innate immunity. However, the mechanism of mA modification in chicken cecum inoculation remains unclear. METHODS: Here, we characterized the cecal mA modification landscape of chicken in the -resistant (R) and susceptible (S) groups using methylated RNA immunoprecipitation sequencing and RNA sequencing (RNA-seq), and further conducted the in vitro inflammatory model based on chicken macrophage-like cell line (HD11) to elucidate the specific mechanism. RESULTS: In the S group, the level of proinflammatory cytokines (IL-8, IL-1β, IL-18, TNF-α, IL-17A) and global RNA methylation were significantly decreased ( < 0.05). A total of 30,427 and 30,367 mA peaks were identified in R and S groups, which were primarily located in 3'UTR and CDS regions. Among these, 514 differential mA peaks (270 hypermethylated peaks and 244 hypomethylated peaks) were identified, which mainly correlated with the regulation of canonical NF-kappaB signal transduction, apoptotic signaling pathway, and MyD88-dependent toll-like receptor signaling pathway. Moreover, we identified 365 differentially expressed genes (DEGs), which were mainly associated with regulation of autophagy, and toll-like receptor 9 signaling pathway, intraciliary transport involved in cilium assembly, positive regulation of mTOR signaling, defense response to bacteria. The correlation analysis revealed that mA methylation level correlated positively with gene expression. Further analysis identified 58 differentially methylated genes (DMGs), and mainly involved in apoptosis, autophagy, Notch signaling pathway and defense response to bacteria, which mainly enriched by DMGs including and . Furthermore, we found that YTHDC2 could involve in regulating the apoptosis and autophagy process of HD11 cells through altering the expression of DMGs including and , which was confirmed by experiments in vitro. CONCLUSION: This result suggested the regulatory role of mA methylation in chicken responds to inoculation. Collectively, the current study characterized the mA modification landscape of chicken cecum and identified YTHDC2 acting key regulator responsible for inoculation.
引言：作为一种食源性共生病原菌，对人类健康和家禽业构成严重威胁。N6-甲基腺苷（m⁶A）mRNA修饰与先天免疫相关。然而，鸡盲肠接种中m⁶A修饰的机制仍不清楚。 方法：在此，我们使用甲基化RNA免疫沉淀测序和RNA测序（RNA-seq）对鸡抗沙门氏菌（R）组和易感（S）组的盲肠m⁶A修饰图谱进行了表征，并基于鸡巨噬细胞样细胞系（HD11）建立了体外炎症模型以阐明具体机制。 结果：在S组中，促炎细胞因子（IL-8、IL-1β、IL-18、TNF-α、IL-17A）水平和整体RNA甲基化水平显著降低（P<0.05）。在R组和S组中分别鉴定出30427个和30367个m⁶A峰，主要位于3'UTR和CDS区域。其中，鉴定出514个差异m⁶A峰（270个高甲基化峰和244个低甲基化峰），主要与经典NF-κB信号转导、凋亡信号通路和MyD88依赖性Toll样受体信号通路的调控相关。此外，我们鉴定出365个差异表达基因（DEGs），主要与自噬调控、Toll样受体9信号通路、参与纤毛组装的纤毛内运输、mTOR信号的正调控、对细菌的防御反应相关。相关性分析表明m⁶A甲基化水平与基因表达呈正相关。进一步分析鉴定出58个差异甲基化基因（DMGs），主要涉及凋亡、自噬、Notch信号通路和对细菌的防御反应，主要由包括[具体基因1]和[具体基因2]在内的DMGs富集。此外，我们发现YTHDC2可通过改变包括[具体基因1]和[具体基因2]在内的DMGs的表达参与调节HD11细胞的凋亡和自噬过程，这在体外实验中得到了证实。 结论：该结果表明m⁶A甲基化在鸡对沙门氏菌接种反应中的调节作用。总体而言，当前研究表征了鸡盲肠的m⁶A修饰图谱，并鉴定出YTHDC2是负责沙门氏菌接种的关键调节因子。
Front Cardiovasc Med. 2025-6-18
Poult Sci. 2025-5-26
Proc Natl Acad Sci U S A. 2024-10-22
J Genet Genomics. 2024-12
Front Microbiol. 2023-12-18
Zotero 插件
