{"title":"METTL3 通过 IGF2BP1 和 YTHDF1 介导的 m6A 修饰提高 YAP 活性，从而促进人类牙周韧带细胞的成骨分化。","authors":"Xuefei Sun, Xiujiao Meng, Yu Piao, Shaojie Dong, Qianqian Dong","doi":"10.1111/jre.13297","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Aims</h3>\n \n <p>N6-Methyladenosine (m<sup>6</sup>A) has been confirmed to play a dynamic role in osteoporosis and bone metabolism. However, whether m<sup>6</sup>A is involved in the osteogenic differentiation of human periodontal ligament cells (hPDLCs) remains unclear. The present study aimed to verify the role of methyltransferase-like 3 (METTL3)-mediated m<sup>6</sup>A modification in the osteogenic differentiation of hPDLCs.</p>\n </section>\n \n <section>\n \n <h3> Methods</h3>\n \n <p>The METTL3, Runx2, Osx, and YAP mRNA expression was determined by qPCR. METTL3, RUNX2, OSX, YTHDF1, YAP, IGF2BP1, and eIF3a protein expression was measured by Western blotting and immunofluorescence assays. The levels of m<sup>6</sup>A modification were evaluated by methylated RNA immunoprecipitation (MeRIP) and dot blot analyses. MeRIP-seq and RNA-seq were used to screen potential candidate genes. Nucleic acid and protein interactions were detected by immunoprecipitation. Alizarin red staining was used to evaluate the osteogenic differentiation of hPDLCs. Gene transcription and promoter activities were assessed by luciferase reporter assays (<i>n</i> ≥ 3).</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>The expression of METTL3 and m<sup>6</sup>A modifications increased synchronously with the osteogenic differentiation of hPDLCs (<i>p</i> = .0016). YAP was a candidate gene identified by MeRIP-seq and RNA-seq, and its mRNA and protein expression levels were simultaneously increased. METTL3 increased the m<sup>6</sup>A methylated IGF2BP1-mediated stability of YAP mRNA (<i>p</i> = .0037), which in turn promoted osteogenic differentiation (<i>p</i> = .0147). Furthermore, METTL3 increased the translation efficiency of YAP by recruiting YTHDF1 and eIF3a to the translation initiation complex (<i>p</i> = .0154), thereby promoting the osteogenic differentiation of hPDLCs (<i>p</i> = .0012).</p>\n </section>\n \n <section>\n \n <h3> Conclusion</h3>\n \n <p>Our study revealed that METTL3-initiated m<sup>6</sup>A mRNA methylation promotes osteogenic differentiation of hPDLCs by increasing IGF2BP1-mediated YAP mRNA stability and recruiting YTHDF1 and eIF3a to the translation initiation complex to increase YAP mRNA translation. Our findings reveal the mechanism of METTL3-mediated m<sup>6</sup>A modification during hPDLC osteogenesis, providing a potential therapeutic target for periodontitis and alveolar bone defects.</p>\n </section>\n </div>","PeriodicalId":16715,"journal":{"name":"Journal of periodontal research","volume":"59 5","pages":"1017-1030"},"PeriodicalIF":3.4000,"publicationDate":"2024-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"METTL3 promotes the osteogenic differentiation of human periodontal ligament cells by increasing YAP activity via IGF2BP1 and YTHDF1-mediated m6A modification\",\"authors\":\"Xuefei Sun, Xiujiao Meng, Yu Piao, Shaojie Dong, Qianqian Dong\",\"doi\":\"10.1111/jre.13297\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n \\n <section>\\n \\n <h3> Aims</h3>\\n \\n <p>N6-Methyladenosine (m<sup>6</sup>A) has been confirmed to play a dynamic role in osteoporosis and bone metabolism. However, whether m<sup>6</sup>A is involved in the osteogenic differentiation of human periodontal ligament cells (hPDLCs) remains unclear. The present study aimed to verify the role of methyltransferase-like 3 (METTL3)-mediated m<sup>6</sup>A modification in the osteogenic differentiation of hPDLCs.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Methods</h3>\\n \\n <p>The METTL3, Runx2, Osx, and YAP mRNA expression was determined by qPCR. METTL3, RUNX2, OSX, YTHDF1, YAP, IGF2BP1, and eIF3a protein expression was measured by Western blotting and immunofluorescence assays. The levels of m<sup>6</sup>A modification were evaluated by methylated RNA immunoprecipitation (MeRIP) and dot blot analyses. MeRIP-seq and RNA-seq were used to screen potential candidate genes. Nucleic acid and protein interactions were detected by immunoprecipitation. Alizarin red staining was used to evaluate the osteogenic differentiation of hPDLCs. Gene transcription and promoter activities were assessed by luciferase reporter assays (<i>n</i> ≥ 3).</p>\\n </section>\\n \\n <section>\\n \\n <h3> Results</h3>\\n \\n <p>The expression of METTL3 and m<sup>6</sup>A modifications increased synchronously with the osteogenic differentiation of hPDLCs (<i>p</i> = .0016). YAP was a candidate gene identified by MeRIP-seq and RNA-seq, and its mRNA and protein expression levels were simultaneously increased. METTL3 increased the m<sup>6</sup>A methylated IGF2BP1-mediated stability of YAP mRNA (<i>p</i> = .0037), which in turn promoted osteogenic differentiation (<i>p</i> = .0147). Furthermore, METTL3 increased the translation efficiency of YAP by recruiting YTHDF1 and eIF3a to the translation initiation complex (<i>p</i> = .0154), thereby promoting the osteogenic differentiation of hPDLCs (<i>p</i> = .0012).</p>\\n </section>\\n \\n <section>\\n \\n <h3> Conclusion</h3>\\n \\n <p>Our study revealed that METTL3-initiated m<sup>6</sup>A mRNA methylation promotes osteogenic differentiation of hPDLCs by increasing IGF2BP1-mediated YAP mRNA stability and recruiting YTHDF1 and eIF3a to the translation initiation complex to increase YAP mRNA translation. Our findings reveal the mechanism of METTL3-mediated m<sup>6</sup>A modification during hPDLC osteogenesis, providing a potential therapeutic target for periodontitis and alveolar bone defects.</p>\\n </section>\\n </div>\",\"PeriodicalId\":16715,\"journal\":{\"name\":\"Journal of periodontal research\",\"volume\":\"59 5\",\"pages\":\"1017-1030\"},\"PeriodicalIF\":3.4000,\"publicationDate\":\"2024-06-04\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of periodontal research\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1111/jre.13297\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"DENTISTRY, ORAL SURGERY & MEDICINE\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of periodontal research","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/jre.13297","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"DENTISTRY, ORAL SURGERY & MEDICINE","Score":null,"Total":0}
METTL3 promotes the osteogenic differentiation of human periodontal ligament cells by increasing YAP activity via IGF2BP1 and YTHDF1-mediated m6A modification
Aims
N6-Methyladenosine (m6A) has been confirmed to play a dynamic role in osteoporosis and bone metabolism. However, whether m6A is involved in the osteogenic differentiation of human periodontal ligament cells (hPDLCs) remains unclear. The present study aimed to verify the role of methyltransferase-like 3 (METTL3)-mediated m6A modification in the osteogenic differentiation of hPDLCs.
Methods
The METTL3, Runx2, Osx, and YAP mRNA expression was determined by qPCR. METTL3, RUNX2, OSX, YTHDF1, YAP, IGF2BP1, and eIF3a protein expression was measured by Western blotting and immunofluorescence assays. The levels of m6A modification were evaluated by methylated RNA immunoprecipitation (MeRIP) and dot blot analyses. MeRIP-seq and RNA-seq were used to screen potential candidate genes. Nucleic acid and protein interactions were detected by immunoprecipitation. Alizarin red staining was used to evaluate the osteogenic differentiation of hPDLCs. Gene transcription and promoter activities were assessed by luciferase reporter assays (n ≥ 3).
Results
The expression of METTL3 and m6A modifications increased synchronously with the osteogenic differentiation of hPDLCs (p = .0016). YAP was a candidate gene identified by MeRIP-seq and RNA-seq, and its mRNA and protein expression levels were simultaneously increased. METTL3 increased the m6A methylated IGF2BP1-mediated stability of YAP mRNA (p = .0037), which in turn promoted osteogenic differentiation (p = .0147). Furthermore, METTL3 increased the translation efficiency of YAP by recruiting YTHDF1 and eIF3a to the translation initiation complex (p = .0154), thereby promoting the osteogenic differentiation of hPDLCs (p = .0012).
Conclusion
Our study revealed that METTL3-initiated m6A mRNA methylation promotes osteogenic differentiation of hPDLCs by increasing IGF2BP1-mediated YAP mRNA stability and recruiting YTHDF1 and eIF3a to the translation initiation complex to increase YAP mRNA translation. Our findings reveal the mechanism of METTL3-mediated m6A modification during hPDLC osteogenesis, providing a potential therapeutic target for periodontitis and alveolar bone defects.
期刊介绍：
The Journal of Periodontal Research is an international research periodical the purpose of which is to publish original clinical and basic investigations and review articles concerned with every aspect of periodontology and related sciences. Brief communications (1-3 journal pages) are also accepted and a special effort is made to ensure their rapid publication. Reports of scientific meetings in periodontology and related fields are also published.
One volume of six issues is published annually.
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