PDF(Pubmed)
Abstract：
UNASSIGNED: Middle ear cholesteatoma is a chronic middle ear disease characterized by severe hearing loss and adjacent bone erosion, resulting in numerous complications. This study sought to identify pathways involved in N6-methyladenosine (m6A) modification of circRNA in middle ear cholesteatoma.
UNASSIGNED: A m6A circRNA epitranscriptomic microarray analysis was performed in middle ear cholesteatoma tissues (n = 5) and normal post-auricular skin samples (n = 5). Bioinformatics analyses subsequently explored the biological functions (Gene Ontology, GO) and signaling pathways (Kyoto Encyclopedia of Genes and Genomes, KEGG) underlying middle ear cholesteatoma pathogenesis. Methylated RNA immunoprecipitation qPCR (MeRIP-qPCR) was performed to verify the presence of circRNAs with m6A modifications in middle ear cholesteatoma and normal skin samples.
UNASSIGNED: Microarray analysis identified 3,755 circRNAs as significantly differentially modified by m6A methylation in middle ear cholesteatoma compared with the normal post-auricular skin. Among these, 3,742 were hypermethylated (FC ≥ 2, FDR < 0.05) and 13 were hypomethylated (FC ≤ 1/2, FDR < 0.05). GO analysis terms with the highest enrichment score were localization, cytoplasm, and ATP-dependent activity for biological processes, cellular components, and molecular functions respectively. Of the eight hypermethylated circRNA pathways, RNA degradation pathway has the highest enrichment score. Peroxisome Proliferator-Activated Receptor (PPAR) signaling pathway was hypomethylated. To validate the microarray analysis, we conducted MeRIP-qPCR to assess the methylation levels of five specific m6A-modified circRNAs: hsa_circRNA_061554, hsa_circRNA_001454, hsa_circRNA_031526, hsa_circRNA_100833, and hsa_circRNA_022382. The validation was highly consistent with the findings from the microarray analysis.
UNASSIGNED: Our study firstly presents m6A modification patterns of circRNAs in middle ear cholesteatoma. This finding suggests a direction for circRNA m6A modification research in the etiology of cholesteatoma and provides potential therapeutic targets for the treatment of middle ear cholesteatoma.
UNASSIGNED: A m6A circRNA epitranscriptomic microarray analysis was performed in middle ear cholesteatoma tissues (n = 5) and normal post-auricular skin samples (n = 5). Bioinformatics analyses subsequently explored the biological functions (Gene Ontology, GO) and signaling pathways (Kyoto Encyclopedia of Genes and Genomes, KEGG) underlying middle ear cholesteatoma pathogenesis. Methylated RNA immunoprecipitation qPCR (MeRIP-qPCR) was performed to verify the presence of circRNAs with m6A modifications in middle ear cholesteatoma and normal skin samples.
UNASSIGNED: Microarray analysis identified 3,755 circRNAs as significantly differentially modified by m6A methylation in middle ear cholesteatoma compared with the normal post-auricular skin. Among these, 3,742 were hypermethylated (FC ≥ 2, FDR < 0.05) and 13 were hypomethylated (FC ≤ 1/2, FDR < 0.05). GO analysis terms with the highest enrichment score were localization, cytoplasm, and ATP-dependent activity for biological processes, cellular components, and molecular functions respectively. Of the eight hypermethylated circRNA pathways, RNA degradation pathway has the highest enrichment score. Peroxisome Proliferator-Activated Receptor (PPAR) signaling pathway was hypomethylated. To validate the microarray analysis, we conducted MeRIP-qPCR to assess the methylation levels of five specific m6A-modified circRNAs: hsa_circRNA_061554, hsa_circRNA_001454, hsa_circRNA_031526, hsa_circRNA_100833, and hsa_circRNA_022382. The validation was highly consistent with the findings from the microarray analysis.
UNASSIGNED: Our study firstly presents m6A modification patterns of circRNAs in middle ear cholesteatoma. This finding suggests a direction for circRNA m6A modification research in the etiology of cholesteatoma and provides potential therapeutic targets for the treatment of middle ear cholesteatoma.
摘要：
■中耳胆脂瘤是一种慢性中耳疾病，其特征是严重的听力损失和邻近的骨侵蚀，导致许多并发症。这项研究旨在鉴定中耳胆脂瘤中circRNA的N6-甲基腺苷（m6A）修饰的途径。
■在中耳胆脂瘤组织(n=5)和正常耳后皮肤样本(n=5)中进行m6AcircircRNA表位基因组微阵列分析。生物信息学分析随后探索了生物学功能(基因本体论，GO）和信号通路（京都基因和基因组百科全书，KEGG）潜在的中耳胆脂瘤发病机制。进行甲基化RNA免疫沉淀qPCR(MeRIP-qPCR)以验证在中耳胆脂瘤和正常皮肤样品中存在具有m6A修饰的circRNA。
■微阵列分析发现，与正常耳后皮肤相比，中耳胆脂瘤中的3,755个circRNAs被m6A甲基化显著差异修饰。其中,3,742例高甲基化（FC≥2，FDR<0.05），13例低甲基化（FC≤1/2，FDR<0.05）。富集得分最高的GO分析术语是本地化，细胞质，和生物过程的ATP依赖性活性，细胞成分，和分子功能分别。在八种高甲基化的circRNA途径中，RNA降解途径具有最高的富集得分。过氧化物酶体增殖物激活受体(PPAR)信号通路被低甲基化。为了验证微阵列分析，我们进行了MeRIP-qPCR，以评估5种特定的m6A修饰的circRNAs的甲基化水平：hsa_circRNA_061554，hsa_circRNA_001454，hsa_circRNA_031526，hsa_circRNA_100833和hsa_circRNA_022382。验证与来自微阵列分析的发现高度一致。
■我们的研究首次提出了中耳胆脂瘤中circRNAs的m6A修饰模式。这一发现为胆脂瘤病因学中的circRNAm6A修饰研究提供了方向，并为中耳胆脂瘤的治疗提供了潜在的治疗靶点。
■在中耳胆脂瘤组织(n=5)和正常耳后皮肤样本(n=5)中进行m6AcircircRNA表位基因组微阵列分析。生物信息学分析随后探索了生物学功能(基因本体论，GO）和信号通路（京都基因和基因组百科全书，KEGG）潜在的中耳胆脂瘤发病机制。进行甲基化RNA免疫沉淀qPCR(MeRIP-qPCR)以验证在中耳胆脂瘤和正常皮肤样品中存在具有m6A修饰的circRNA。
■微阵列分析发现，与正常耳后皮肤相比，中耳胆脂瘤中的3,755个circRNAs被m6A甲基化显著差异修饰。其中,3,742例高甲基化（FC≥2，FDR<0.05），13例低甲基化（FC≤1/2，FDR<0.05）。富集得分最高的GO分析术语是本地化，细胞质，和生物过程的ATP依赖性活性，细胞成分，和分子功能分别。在八种高甲基化的circRNA途径中，RNA降解途径具有最高的富集得分。过氧化物酶体增殖物激活受体(PPAR)信号通路被低甲基化。为了验证微阵列分析，我们进行了MeRIP-qPCR，以评估5种特定的m6A修饰的circRNAs的甲基化水平：hsa_circRNA_061554，hsa_circRNA_001454，hsa_circRNA_031526，hsa_circRNA_100833和hsa_circRNA_022382。验证与来自微阵列分析的发现高度一致。
■我们的研究首次提出了中耳胆脂瘤中circRNAs的m6A修饰模式。这一发现为胆脂瘤病因学中的circRNAm6A修饰研究提供了方向，并为中耳胆脂瘤的治疗提供了潜在的治疗靶点。
