{"title":"高通量测序揭示N6-甲基腺苷修饰的lncRNA作为肝纤维化小鼠的潜在生物标志物。","authors":"Furong Wu, Shengyu Zhang, Chang Fan, Shaopeng Huang, Hui Jiang, Jiafu Zhang","doi":"10.2174/1566523223666230606151013","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>N<sup>6</sup>-methyladenosine (m<sup>6</sup>A) is the most frequent internal modification in eukaryotic RNA. Long noncoding RNAs (lncRNAs) are a new type of noncoding regulatory molecule with multiple cellular functions. Both are closely related to the occurrence and development of liver fibrosis (LF). However, the role of m<sup>6</sup>A-methylated lncRNAs in the progression of LF remains largely unknown.</p><p><strong>Methods: </strong>In this study, HE and Masson staining were used to observe pathological changes in the liver, m<sup>6</sup>A-modified RNA immunoprecipitation sequencing (m<sup>6</sup>A-seq) was performed to systematically evaluate the m<sup>6</sup>A modification level of lncRNAs in LF mice, meRIP-qPCR and RT-qPCR were used to detect the m<sup>6</sup>A methylation level and RNA expression level of the target lncRNAs.</p><p><strong>Results: </strong>A total of 415 m<sup>6</sup>A peaks were detected in 313 lncRNAs in liver fibrosis tissues. There were 98 significantly different m<sup>6</sup>A peaks in LF, which were located on 84 lncRNAs, of which 45.2% of the lncRNA length was between 200-400 bp. At the same time, the first three chromosomes of these methylated lncRNAs were chromosomes 7, 5 and 1. RNA sequencing identified 154 differentially expressed lncRNAs in LF. The joint analysis of m<sup>6</sup>A-seq and RNA-seq found that there were three lncRNAs with significant changes in m<sup>6</sup>A methylation and RNA expression levels: lncRNA H19, lncRNA Gm16023 and lncRNA Gm17586. Subsequently, the verification results showed that the m<sup>6</sup>A methylation levels of lncRNA H19 and lncRNA Gm17586 were significantly increased, while that of lncRNA Gm16023 was significantly decreased, and the RNA expression of three lncRNAs was significantly decreased. Through the establishment of a lncRNA-miRNA-mRNA regulatory network, the possible regulatory relationships of lncRNA H19, lncRNA Gm16023 and lncRNA Gm17586 in LF were revealed.</p><p><strong>Conclusion: </strong>This study revealed the unique m<sup>6</sup>A methylation pattern of lncRNAs in LF mice, suggesting that the m<sup>6</sup>A methylation modification of lncRNAs is related to the occurrence and development of LF.</p>","PeriodicalId":10798,"journal":{"name":"Current gene therapy","volume":" ","pages":"371-390"},"PeriodicalIF":3.8000,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"High-Throughput Sequencing Reveals N<sup>6</sup>-Methyladenosine-modified LncRNAs as Potential Biomarkers in Mice with Liver Fibrosis.\",\"authors\":\"Furong Wu, Shengyu Zhang, Chang Fan, Shaopeng Huang, Hui Jiang, Jiafu Zhang\",\"doi\":\"10.2174/1566523223666230606151013\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>N<sup>6</sup>-methyladenosine (m<sup>6</sup>A) is the most frequent internal modification in eukaryotic RNA. Long noncoding RNAs (lncRNAs) are a new type of noncoding regulatory molecule with multiple cellular functions. Both are closely related to the occurrence and development of liver fibrosis (LF). However, the role of m<sup>6</sup>A-methylated lncRNAs in the progression of LF remains largely unknown.</p><p><strong>Methods: </strong>In this study, HE and Masson staining were used to observe pathological changes in the liver, m<sup>6</sup>A-modified RNA immunoprecipitation sequencing (m<sup>6</sup>A-seq) was performed to systematically evaluate the m<sup>6</sup>A modification level of lncRNAs in LF mice, meRIP-qPCR and RT-qPCR were used to detect the m<sup>6</sup>A methylation level and RNA expression level of the target lncRNAs.</p><p><strong>Results: </strong>A total of 415 m<sup>6</sup>A peaks were detected in 313 lncRNAs in liver fibrosis tissues. There were 98 significantly different m<sup>6</sup>A peaks in LF, which were located on 84 lncRNAs, of which 45.2% of the lncRNA length was between 200-400 bp. At the same time, the first three chromosomes of these methylated lncRNAs were chromosomes 7, 5 and 1. RNA sequencing identified 154 differentially expressed lncRNAs in LF. The joint analysis of m<sup>6</sup>A-seq and RNA-seq found that there were three lncRNAs with significant changes in m<sup>6</sup>A methylation and RNA expression levels: lncRNA H19, lncRNA Gm16023 and lncRNA Gm17586. Subsequently, the verification results showed that the m<sup>6</sup>A methylation levels of lncRNA H19 and lncRNA Gm17586 were significantly increased, while that of lncRNA Gm16023 was significantly decreased, and the RNA expression of three lncRNAs was significantly decreased. Through the establishment of a lncRNA-miRNA-mRNA regulatory network, the possible regulatory relationships of lncRNA H19, lncRNA Gm16023 and lncRNA Gm17586 in LF were revealed.</p><p><strong>Conclusion: </strong>This study revealed the unique m<sup>6</sup>A methylation pattern of lncRNAs in LF mice, suggesting that the m<sup>6</sup>A methylation modification of lncRNAs is related to the occurrence and development of LF.</p>\",\"PeriodicalId\":10798,\"journal\":{\"name\":\"Current gene therapy\",\"volume\":\" \",\"pages\":\"371-390\"},\"PeriodicalIF\":3.8000,\"publicationDate\":\"2023-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current gene therapy\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.2174/1566523223666230606151013\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"GENETICS & HEREDITY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current gene therapy","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.2174/1566523223666230606151013","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"GENETICS & HEREDITY","Score":null,"Total":0}
High-Throughput Sequencing Reveals N6-Methyladenosine-modified LncRNAs as Potential Biomarkers in Mice with Liver Fibrosis.
Background: N6-methyladenosine (m6A) is the most frequent internal modification in eukaryotic RNA. Long noncoding RNAs (lncRNAs) are a new type of noncoding regulatory molecule with multiple cellular functions. Both are closely related to the occurrence and development of liver fibrosis (LF). However, the role of m6A-methylated lncRNAs in the progression of LF remains largely unknown.
Methods: In this study, HE and Masson staining were used to observe pathological changes in the liver, m6A-modified RNA immunoprecipitation sequencing (m6A-seq) was performed to systematically evaluate the m6A modification level of lncRNAs in LF mice, meRIP-qPCR and RT-qPCR were used to detect the m6A methylation level and RNA expression level of the target lncRNAs.
Results: A total of 415 m6A peaks were detected in 313 lncRNAs in liver fibrosis tissues. There were 98 significantly different m6A peaks in LF, which were located on 84 lncRNAs, of which 45.2% of the lncRNA length was between 200-400 bp. At the same time, the first three chromosomes of these methylated lncRNAs were chromosomes 7, 5 and 1. RNA sequencing identified 154 differentially expressed lncRNAs in LF. The joint analysis of m6A-seq and RNA-seq found that there were three lncRNAs with significant changes in m6A methylation and RNA expression levels: lncRNA H19, lncRNA Gm16023 and lncRNA Gm17586. Subsequently, the verification results showed that the m6A methylation levels of lncRNA H19 and lncRNA Gm17586 were significantly increased, while that of lncRNA Gm16023 was significantly decreased, and the RNA expression of three lncRNAs was significantly decreased. Through the establishment of a lncRNA-miRNA-mRNA regulatory network, the possible regulatory relationships of lncRNA H19, lncRNA Gm16023 and lncRNA Gm17586 in LF were revealed.
Conclusion: This study revealed the unique m6A methylation pattern of lncRNAs in LF mice, suggesting that the m6A methylation modification of lncRNAs is related to the occurrence and development of LF.
期刊介绍：
Current Gene Therapy is a bi-monthly peer-reviewed journal aimed at academic and industrial scientists with an interest in major topics concerning basic research and clinical applications of gene and cell therapy of diseases. Cell therapy manuscripts can also include application in diseases when cells have been genetically modified. Current Gene Therapy publishes full-length/mini reviews and original research on the latest developments in gene transfer and gene expression analysis, vector development, cellular genetic engineering, animal models and human clinical applications of gene and cell therapy for the treatment of diseases.
Current Gene Therapy publishes reviews and original research containing experimental data on gene and cell therapy. The journal also includes manuscripts on technological advances, ethical and regulatory considerations of gene and cell therapy. Reviews should provide the reader with a comprehensive assessment of any area of experimental biology applied to molecular medicine that is not only of significance within a particular field of gene therapy and cell therapy but also of interest to investigators in other fields. Authors are encouraged to provide their own assessment and vision for future advances. Reviews are also welcome on late breaking discoveries on which substantial literature has not yet been amassed. Such reviews provide a forum for sharply focused topics of recent experimental investigations in gene therapy primarily to make these results accessible to both clinical and basic researchers. Manuscripts containing experimental data should be original data, not previously published.
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