PMID- 38358895
OWN - NLM
STAT- MEDLINE
DCOM- 20240313
LR  - 20240329
IS  - 1573-4935 (Electronic)
IS  - 0144-8463 (Print)
IS  - 0144-8463 (Linking)
VI  - 44
IP  - 3
DP  - 2024 Mar 29
TI  - METTL3 promotes osteogenic differentiation of human umbilical cord mesenchymal 
      stem cells by up-regulating m6A modification of circCTTN.
LID - 10.1042/BSR20231186 [doi]
LID - BSR20231186
AB  - BACKGROUND: Human umbilical cord mesenchymal stem cells (hUCMSCs) are promising 
      seed cells in bone tissue engineering. circRNA and N6-methyladenosine (m6A) RNA 
      methylation play important roles in osteogenic differentiation. Here, we 
      investigated the potential relevance of a critical circRNA, hsa_circ_0003376 
      (circCTTN), and methyltransferase-like 3 (METTL3) in osteogenic differentiation 
      of hUCMSCs. METHODS: Expression of circCTTN after hUCMSC osteogenic induction was 
      detected by qRT-PCR. Three databases (RMBase v2.0, BERMP, and SRAMP) were used to 
      predict m6A sites of circCTTN. RNA was enriched by methylated RNA 
      immunoprecipitation (MeRIP), followed by quantitative real-time polymerase chain 
      reaction to detect m6A level of circCTTN after METTL3 overexpression and 
      osteogenic induction. RNA pull-down, Western blotting, and protein mass 
      spectrometry were performed to investigate the potential mechanisms by which 
      METTL3 promoted m6A modification of circCTTN. Bioinformatic analyses based on 
      database (STRING) search and co-immunoprecipitation were used to analyze the 
      proteins that interacted with METTL3. RESULTS: Overexpression of METTL3 promoted 
      osteogenic differentiation of hUCMSCs and increased m6A level of circCTTN. Two 
      potential m6A modification sites of circCTTN were predicted. No direct 
      interaction between METTL3 and circCTTN was observed. Thirty-one proteins were 
      pulled down by probes specific for circCTTN, including NOP2, and two m6A reading 
      proteins, EIF3A and SND1. Bioinformatics analysis and co-immunoprecipitation 
      showed that METTL3 interacted with EIF3A indirectly through NOP2. CONCLUSIONS: 
      METTL3 promotes the osteogenic differentiation of hUCMSCs by increasing the m6A 
      level of circCTTN. However, METTL3 does not bind directly to circCTTN. METTL3 
      interacts with circCTTN indirectly through NOP2 and EIF3A.
CI  - (c) 2024 The Author(s).
FAU - Chen, Shujiang
AU  - Chen S
AD  - State Key Laboratory of Oral Diseases and National Clinical Research Center for 
      Oral Diseases, West China school of Stomatology, Sichuan University, Chengdu, 
      Sichuan, China.
AD  - Department of Prosthodontics, West China Hospital of Stomatology, Sichuan 
      University, Chengdu, Sichuan, China.
FAU - Duan, Xiaoqiong
AU  - Duan X
AD  - Institute of Blood Transfusion, Chinese Academy of Medical Sciences and Peking 
      Union Medical College, Chengdu, Sichuan, China.
FAU - He, Yanjin
AU  - He Y
AD  - State Key Laboratory of Oral Diseases and National Clinical Research Center for 
      Oral Diseases, West China school of Stomatology, Sichuan University, Chengdu, 
      Sichuan, China.
AD  - Department of Prosthodontics, West China Hospital of Stomatology, Sichuan 
      University, Chengdu, Sichuan, China.
FAU - Chen, Wenchuan
AU  - Chen W
AD  - State Key Laboratory of Oral Diseases and National Clinical Research Center for 
      Oral Diseases, West China school of Stomatology, Sichuan University, Chengdu, 
      Sichuan, China.
AD  - Department of Prosthodontics, West China Hospital of Stomatology, Sichuan 
      University, Chengdu, Sichuan, China.
LA  - eng
GR  - 82270957/the National Natural Science Foundation of China/
GR  - 2020YJ0228/Science and Technology Department of Sichuan Province (SPDST)/
PT  - Journal Article
PL  - England
TA  - Biosci Rep
JT  - Bioscience reports
JID - 8102797
RN  - EC 3.1.- (Endonucleases)
RN  - EC 2.1.1.- (Methyltransferases)
RN  - EC 2.1.1.62 (METTL3 protein, human)
RN  - CLE6G00625 (N-methyladenosine)
RN  - 63231-63-0 (RNA)
RN  - 0 (RNA, Circular)
RN  - EC 3.1.- (SND1 protein, human)
SB  - IM
MH  - Humans
MH  - Endonucleases
MH  - *Mesenchymal Stem Cells
MH  - *Methyltransferases/genetics
MH  - *Osteogenesis/genetics
MH  - RNA
MH  - RNA, Circular/genetics/metabolism
MH  - Umbilical Cord
PMC - PMC10932744
OTO - NOTNLM
OT  - METTL3
OT  - circCTTN
OT  - human umbilical cord mesenchymal stromal cells
OT  - m6A
OT  - osteogenic differentiation
COIS- The authors declare that there are no competing interests associated with the 
      manuscript.
EDAT- 2024/02/15 18:42
MHDA- 2024/03/13 06:46
PMCR- 2024/03/12
CRDT- 2024/02/15 12:42
PHST- 2023/07/28 00:00 [received]
PHST- 2024/02/10 00:00 [revised]
PHST- 2024/02/13 00:00 [accepted]
PHST- 2024/03/13 06:46 [medline]
PHST- 2024/02/15 18:42 [pubmed]
PHST- 2024/02/15 12:42 [entrez]
PHST- 2024/03/12 00:00 [pmc-release]
AID - 234063 [pii]
AID - BSR20231186 [pii]
AID - 10.1042/BSR20231186 [doi]
PST - ppublish
SO  - Biosci Rep. 2024 Mar 29;44(3):BSR20231186. doi: 10.1042/BSR20231186.