机构：[1]Department of Obstetrics, Affiliated Xiaoshan Hospital, Hangzhou Normal University, Hangzhou, Zhejiang, China.[2]Department of Obstetrics and Gynecology, The First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan, China[3]Department of Obstetrics and Gynecology, Department of Fetal Medicine and Prenatal Diagnosis, Key Laboratory for Major Obstetric Diseases of Guangdong Province, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
Objective: N6-methyladenosine (m6A) is a common post-transcriptional
modification of messenger RNAs (mRNAs) and long non-coding RNAs
(lncRNAs). However, m6A-modified lncRNAs are still largely unexplored. This
study aimed to investigate differentially m6A-modified lncRNAs and genes
involved in neural tube defect (NTD) development.
Methods: Pregnant Kunming mice (9–10 weeks of age) were treated with
retinoic acid to construct NTD models. m6A levels and methyltransferase-like 3
(METTL3) expression were evaluated in brain tissues of the NTD models.
Methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA
sequencing (RNA-seq) were performed on the NovaSeq platform and
Illumina HiSeq 2,500 platform, respectively. Differentially m6A-methylated
differentially expressed lncRNAs (DElncRNAs) and differentially expressed
genes (DEGs) were identified, followed by GO biological process and KEGG
pathway functional enrichment analyses. Expression levels of several
DElncRNAs and DEGs were evaluated by quantitative reverse transcriptionpolymerase
chain reaction (qRT-PCR) for validation.
Results:m6A levels and METTL3 expression levels were significantly lower in the
brain tissues of the NTD mouse model than in controls. By integrating MeRIPseq
and RNA-seq data, 13 differentially m6A-methylated DElncRNAs and
170 differentially m6A-methylated DEGs were identified. They were
significantly enriched in the Hippo signaling pathway and mannose-type
O-glycan biosynthesis. The qRT-PCR results confirmed the decreased
expression levels of lncRNAs, such as Mir100hg, Gm19265, Gm10544, and
Malat1, and genes, such as Zfp236, Erc2, and Hmg20a, in the NTD group.
Conclusion: METTL3-mediated m6A modifications may be involved in NTD
development. In particular, decreased expression levels of Mir100hg, Gm19265,
Gm10544, Malat1, Zfp236, Erc2, and Hmg20a may contribute to the
development of NTD.
基金：
This study was supported by the Guangzhou Science and
Technology Program (No. 202102010129), Yunnan Province
Science and Technology Program (No. 202101AY070001-121)
and the Hangzhou Science and Technology Program (No.
20211231Y121).
第一作者机构：[1]Department of Obstetrics, Affiliated Xiaoshan Hospital, Hangzhou Normal University, Hangzhou, Zhejiang, China.
共同第一作者：
通讯作者：
推荐引用方式(GB/T 7714)：
Yang Jing,Jing Xu,Luting Zhang,et al.Identifying key m6A-methylated lncRNAs and genes associated with neural tube defects via integrative MeRIP and RNA sequencing analyses[J].Frontiers in genetics.2022,13:974357.doi:10.3389/fgene.2022.974357.
APA：
Yang Jing,Jing Xu,Luting Zhang,Yingting Li&Min Chen.(2022).Identifying key m6A-methylated lncRNAs and genes associated with neural tube defects via integrative MeRIP and RNA sequencing analyses.Frontiers in genetics,13,
MLA：
Yang Jing,et al."Identifying key m6A-methylated lncRNAs and genes associated with neural tube defects via integrative MeRIP and RNA sequencing analyses".Frontiers in genetics 13.(2022):974357
