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Stage-specific requirement for METTL3-dependent m(6)A modification during dental pulp stem cell differentiation

BACKGROUND: N(6)-methyladenosine (m(6)A) is the most prevalent epigenetic modification in eukaryotic messenger RNAs and plays a critical role in cell fate transition. However, it remains to be elucidated how m(6)A marks functionally impact the transcriptional cascades that orchestrate stem cell diff...

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Autores principales: Luo, Haiyun, Liu, Wenjing, Zhou, Yachuan, Zhang, Yanli, Wu, Junrong, Wang, Ruolan, Shao, Longquan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9756505/
https://www.ncbi.nlm.nih.gov/pubmed/36527141
http://dx.doi.org/10.1186/s12967-022-03814-9
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author Luo, Haiyun
Liu, Wenjing
Zhou, Yachuan
Zhang, Yanli
Wu, Junrong
Wang, Ruolan
Shao, Longquan
author_facet Luo, Haiyun
Liu, Wenjing
Zhou, Yachuan
Zhang, Yanli
Wu, Junrong
Wang, Ruolan
Shao, Longquan
author_sort Luo, Haiyun
collection PubMed
description BACKGROUND: N(6)-methyladenosine (m(6)A) is the most prevalent epigenetic modification in eukaryotic messenger RNAs and plays a critical role in cell fate transition. However, it remains to be elucidated how m(6)A marks functionally impact the transcriptional cascades that orchestrate stem cell differentiation. The present study focuses on the biological function and mechanism of m(6)A methylation in dental pulp stem cell (DPSC) differentiation. METHODS: m(6)A RNA immunoprecipitation sequencing was utilized to assess the m(6)A-mRNA landscape during DPSC differentiation. Ectopic transplantation of DPSCs in immunodeficient mice was conducted to verify the in vitro findings. RNA sequencing and m(6)A RNA immunoprecipitation sequencing were combined to identify the candidate targets. RNA immunoprecipitation and RNA/protein stability of Noggin (NOG) were evaluated. The alteration in poly(A) tail was measured by 3′-RACE and poly(A) tail length assays. RESULTS: We characterized a dynamic m(6)A-mRNA landscape during DPSC mineralization with increasing enrichment in the 3′ untranslated region (UTR). Methyltransferase-like 3 (METTL3) was identified as the key m(6)A player, and METTL3 knockdown disrupted functional DPSC differentiation. Moreover, METTL3 overexpression enhanced DPSC mineralization. Increasing m(6)A deposition in the 3′ UTR restricted NOG expression, which is required for DPSC mineralization. This stage-specific m(6)A methylation and destabilization of NOG was suppressed by METTL3 knockdown only in differentiated DPSCs. Furthermore, METTL3 promotes the degradation of m(6)A-tagged NOG by shortening the poly(A) tail length in the differentiated stage. CONCLUSIONS: Our results address an essential role of dynamic m(6)A signaling in the temporal control of DPSC differentiation and provide new insight into epitranscriptomic mechanisms in stem cell-based therapy. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12967-022-03814-9.
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spelling pubmed-97565052022-12-17 Stage-specific requirement for METTL3-dependent m(6)A modification during dental pulp stem cell differentiation Luo, Haiyun Liu, Wenjing Zhou, Yachuan Zhang, Yanli Wu, Junrong Wang, Ruolan Shao, Longquan J Transl Med Research BACKGROUND: N(6)-methyladenosine (m(6)A) is the most prevalent epigenetic modification in eukaryotic messenger RNAs and plays a critical role in cell fate transition. However, it remains to be elucidated how m(6)A marks functionally impact the transcriptional cascades that orchestrate stem cell differentiation. The present study focuses on the biological function and mechanism of m(6)A methylation in dental pulp stem cell (DPSC) differentiation. METHODS: m(6)A RNA immunoprecipitation sequencing was utilized to assess the m(6)A-mRNA landscape during DPSC differentiation. Ectopic transplantation of DPSCs in immunodeficient mice was conducted to verify the in vitro findings. RNA sequencing and m(6)A RNA immunoprecipitation sequencing were combined to identify the candidate targets. RNA immunoprecipitation and RNA/protein stability of Noggin (NOG) were evaluated. The alteration in poly(A) tail was measured by 3′-RACE and poly(A) tail length assays. RESULTS: We characterized a dynamic m(6)A-mRNA landscape during DPSC mineralization with increasing enrichment in the 3′ untranslated region (UTR). Methyltransferase-like 3 (METTL3) was identified as the key m(6)A player, and METTL3 knockdown disrupted functional DPSC differentiation. Moreover, METTL3 overexpression enhanced DPSC mineralization. Increasing m(6)A deposition in the 3′ UTR restricted NOG expression, which is required for DPSC mineralization. This stage-specific m(6)A methylation and destabilization of NOG was suppressed by METTL3 knockdown only in differentiated DPSCs. Furthermore, METTL3 promotes the degradation of m(6)A-tagged NOG by shortening the poly(A) tail length in the differentiated stage. CONCLUSIONS: Our results address an essential role of dynamic m(6)A signaling in the temporal control of DPSC differentiation and provide new insight into epitranscriptomic mechanisms in stem cell-based therapy. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12967-022-03814-9. BioMed Central 2022-12-16 /pmc/articles/PMC9756505/ /pubmed/36527141 http://dx.doi.org/10.1186/s12967-022-03814-9 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Luo, Haiyun
Liu, Wenjing
Zhou, Yachuan
Zhang, Yanli
Wu, Junrong
Wang, Ruolan
Shao, Longquan
Stage-specific requirement for METTL3-dependent m(6)A modification during dental pulp stem cell differentiation
title Stage-specific requirement for METTL3-dependent m(6)A modification during dental pulp stem cell differentiation
title_full Stage-specific requirement for METTL3-dependent m(6)A modification during dental pulp stem cell differentiation
title_fullStr Stage-specific requirement for METTL3-dependent m(6)A modification during dental pulp stem cell differentiation
title_full_unstemmed Stage-specific requirement for METTL3-dependent m(6)A modification during dental pulp stem cell differentiation
title_short Stage-specific requirement for METTL3-dependent m(6)A modification during dental pulp stem cell differentiation
title_sort stage-specific requirement for mettl3-dependent m(6)a modification during dental pulp stem cell differentiation
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9756505/
https://www.ncbi.nlm.nih.gov/pubmed/36527141
http://dx.doi.org/10.1186/s12967-022-03814-9
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