YTHDC1 修饰的 hsa_circ_0102678 通过调控 miR-146a/TRAF6 和 IRAK1 轴促进痤疮丙酸杆菌生物膜诱导的角质形成细胞炎症。

YTHDC1-Modified m6A Methylation of Hsa_circ_0102678 Promotes Keratinocyte Inflammation Induced by Cutibacterium acnes Biofilm through Regulating miR-146a/TRAF6 and IRAK1 Axis.

机构信息

Department of Laser Surgery, Jiangsu Key Laboratory of Molecular Biology for Skin Diseases and STIs, Hospital of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing, China,

Department of Dermatology, The Affiliated Jiangning Hospital with Nanjing Medical University, Nanjing, China.

出版信息

J Innate Immun. 2023;15(1):822-835. doi: 10.1159/000534704. Epub 2023 Oct 30.

DOI:10.1159/000534704

PMID:37903473

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10684258/

Abstract

INTRODUCTION

CircRNAs are closely related to many human diseases; however, their role in acne remains unclear. This study aimed to determine the role of hsa_circ_0102678 in regulating inflammation of acne.

METHODS

First, microarray analysis was performed to study the expression of circRNAs in acne. Subsequently, RNase R digestion assay and fluorescence in situ hybridization assay were utilized to confirm the characteristics of hsa_circ_0102678. Finally, qRT-PCR, Western blotting analysis, immunoprecipitation, luciferase reporter assay, circRNA probe pull-down assay, biotin-labeled miRNA pull-down assay, RNA immunoprecipitation assay, and m6A dot blot assay were utilized to reveal the functional roles of hsa_circ_0102678 on inflammation induced by C. acnes biofilm in human primary keratinocytes.

RESULTS

Our investigations showed that the expression of hsa_circ_0102678 was significantly decreased in acne tissues, and hsa_circ_0102678 was a type of circRNAs, which was mainly localized in the cytoplasm of primary human keratinocytes. Moreover, hsa_circ_0102678 remarkably affected the expression of IL-8, IL-6, and TNF-α, which induced by C. acnes biofilm. Importantly, mechanistic studies indicated that the YTHDC1 could bind directly to hsa_circ_0102678 and promote the export of N6-methyladenosine-modified hsa_circ_0102678 to the cytoplasm. Besides, hsa_circ_0102678 could bind to miR-146a and sponge miR-146a to promote the expression of IRAK1 and TRAF6.

CONCLUSION

Our findings revealed a previously unknown process by which hsa_circ_0102678 promoted keratinocyte inflammation induced by C. acnes biofilm via regulating miR-146a/TRAF6 and IRAK1 axis.

摘要

简介

circRNAs 与许多人类疾病密切相关；然而，其在痤疮中的作用尚不清楚。本研究旨在确定 hsa_circ_0102678 在调节痤疮炎症中的作用。

方法

首先，进行微阵列分析以研究 circRNAs 在痤疮中的表达。随后，利用 RNase R 消化实验和荧光原位杂交实验来验证 hsa_circ_0102678 的特征。最后，利用 qRT-PCR、Western blot 分析、免疫沉淀、荧光素酶报告基因实验、circRNA 探针下拉实验、生物素标记 miRNA 下拉实验、RNA 免疫沉淀实验和 m6A 斑点印迹实验揭示了 hsa_circ_0102678 在 C. acnes 生物膜诱导的人原代角质细胞炎症中的功能作用。

结果

我们的研究表明，hsa_circ_0102678 在痤疮组织中的表达显著降低，hsa_circ_0102678 是一种 circRNAs，主要定位于原代人角质细胞的细胞质中。此外，hsa_circ_0102678 显著影响 C. acnes 生物膜诱导的 IL-8、IL-6 和 TNF-α 的表达。重要的是，机制研究表明，YTHDC1 可以直接结合 hsa_circ_0102678 并促进 N6-甲基腺苷修饰的 hsa_circ_0102678 向细胞质的输出。此外，hsa_circ_0102678 可以与 miR-146a 结合并海绵 miR-146a 以促进 IRAK1 和 TRAF6 的表达。