PMID- 34215719
OWN - NLM
STAT- MEDLINE
DCOM- 20210920
LR  - 20210920
IS  - 2041-4889 (Electronic)
VI  - 12
IP  - 7
DP  - 2021 Jul 2
TI  - Downregulated lncRNA RCPCD promotes differentiation of embryonic stem cells into 
      cardiac pacemaker-like cells by suppressing HCN4 promoter methylation.
PG  - 667
LID - 10.1038/s41419-021-03949-5 [doi]
LID - 667
AB  - Long non-coding RNA (lncRNA) is receiving increasing attention in embryonic stem 
      cells (ESCs) research. However, the roles of lncRNA in the differentiation of 
      ESCs into pacemaker-like cells are still unclear. Therefore, the present study 
      aims to explore the roles and mechanisms of lncRNA in the differentiation of ESCs 
      into pacemaker-like cells. ESCs were cultured and induced differentiation to 
      pacemaker-like cells. RNA sequencing was used to identify the differential 
      expression lncRNAs during the differentiation of ESCs into pacemaker-like cells. 
      Cell morphology observation, flow cytometry, quantitative real-time polymerase 
      chain reaction, western blot, and immunofluorescence were used to detect the 
      differentiation of ESCs into pacemaker-like cells. LncRNA and genes 
      overexpression or knockdown through transfected adenovirus in the differentiation 
      process. The fluorescence in situ hybridization (FISH) detected the lncRNA 
      location in the differentiated ESCs. Luciferase reporter gene assay, 
      methylation-specific PCR, chromatin immunoprecipitation assay, and RNA 
      immunoprecipitation assay were performed to reveal the mechanism of 
      lncRNA-regulating HCN4 expression. Rescue experiments were used to confirm that 
      lncRNA regulates the differentiation of ESCs into pacemaker-like cells through 
      HCN4. We cultured the ESCs and induced the differentiation of ESCs into 
      pacemaker-like cells successfully. The expression of lncRNA RCPCD was 
      significantly decreased in the differentiation of ESCs into pacemaker-like cells. 
      Overexpression of RCPCD inhibited the differentiation of ESCs into pacemaker-like 
      cells. RCPCD inhibited the expression of HCN4 by increasing HCN4 methylation at 
      the promoter region through DNMT1, DNMT2, and DNMT3. RCPCD inhibited the 
      differentiation of ESCs into pacemaker-like cells by inhibiting the expression of 
      HCN4. Our results confirm the roles and mechanism of lncRNA RCPCD in the 
      differentiation of ESCs into pacemaker-like cells, which could pave the path for 
      the development of a cell-based biological pacemaker.
FAU - Zhu, Ye
AU  - Zhu Y
AD  - Clinical Medical College of Yangzhou University, Yangzhou, China. 
      307971331@qq.com.
AD  - Department of Cardiology, Northern Jiangsu People's Hospital, Yangzhou, China. 
      307971331@qq.com.
FAU - You, Jia
AU  - You J
AD  - Department of Internal Medicine, Yangzhou Maternal and Child Health Care 
      Hospital, Yangzhou, Jiangsu, 225001, China.
FAU - Wei, Wei
AU  - Wei W
AD  - Clinical Medical College of Yangzhou University, Yangzhou, China.
AD  - Department of Cardiology, Northern Jiangsu People's Hospital, Yangzhou, China.
FAU - Gu, Jianjun
AU  - Gu J
AD  - Clinical Medical College of Yangzhou University, Yangzhou, China.
AD  - Department of Cardiology, Northern Jiangsu People's Hospital, Yangzhou, China.
FAU - Xu, Chao
AU  - Xu C
AUID- ORCID: 0000-0002-3821-6187
AD  - Department of Biostatistics and Epidemiology, University of Oklahoma Health 
      Science Center, Oklahoma City, OK, 73104, US.
FAU - Gu, Xiang
AU  - Gu X
AD  - Clinical Medical College of Yangzhou University, Yangzhou, China.
AD  - Department of Cardiology, Northern Jiangsu People's Hospital, Yangzhou, China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20210702
PL  - England
TA  - Cell Death Dis
JT  - Cell death & disease
JID - 101524092
RN  - 0 (Hcn4 protein, mouse)
RN  - 0 (Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels)
RN  - 0 (RNA, Long Noncoding)
RN  - EC 2.1.1.37 (DNA (Cytosine-5-)-Methyltransferases)
SB  - IM
MH  - Animals
MH  - *Biological Clocks
MH  - *Cell Differentiation
MH  - Cells, Cultured
MH  - DNA (Cytosine-5-)-Methyltransferases/metabolism
MH  - *DNA Methylation
MH  - Down-Regulation
MH  - Gene Expression Regulation, Developmental
MH  - Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels/*genetics/metabolism
MH  - Mice
MH  - Mouse Embryonic Stem Cells/*metabolism
MH  - Myocytes, Cardiac/*metabolism
MH  - *Promoter Regions, Genetic
MH  - RNA, Long Noncoding/*genetics/metabolism
MH  - Sinoatrial Node/cytology/*metabolism
PMC - PMC8253811
COIS- The authors declare no competing interests.
EDAT- 2021/07/04 06:00
MHDA- 2021/09/21 06:00
PMCR- 2021/07/02
CRDT- 2021/07/03 05:39
PHST- 2020/11/15 00:00 [received]
PHST- 2021/06/16 00:00 [accepted]
PHST- 2021/06/16 00:00 [revised]
PHST- 2021/07/03 05:39 [entrez]
PHST- 2021/07/04 06:00 [pubmed]
PHST- 2021/09/21 06:00 [medline]
PHST- 2021/07/02 00:00 [pmc-release]
AID - 10.1038/s41419-021-03949-5 [pii]
AID - 3949 [pii]
AID - 10.1038/s41419-021-03949-5 [doi]
PST - epublish
SO  - Cell Death Dis. 2021 Jul 2;12(7):667. doi: 10.1038/s41419-021-03949-5.