PMID- 37972802
OWN - NLM
STAT- MEDLINE
DCOM- 20231204
LR  - 20240322
IS  - 1873-3913 (Electronic)
IS  - 0898-6568 (Linking)
VI  - 113
DP  - 2024 Jan
TI  - LncRNA Malat1 regulates iPSC-derived beta-cell differentiation by targeting the 
      miR-15b-5p/Ihh axis.
PG  - 110975
LID - S0898-6568(23)00390-X [pii]
LID - 10.1016/j.cellsig.2023.110975 [doi]
AB  - BACKGROUND: Differentiation of induced pluripotent stem cells (iPSCs)-derived 
      beta-like cells is a novel strategy for treatment of type 1 diabetes. Elucidation of 
      the regulatory mechanisms of long noncoding RNAs (lncRNAs) in beta-like cells 
      derived from iPSCs is important for understanding the development of the pancreas 
      and pancreatic beta-cells and may improve the quality of beta-like cells for stem cell 
      therapy. METHODS: beta-like cells were derived from iPSCs in a three-step protocol. 
      RNA sequencing and bioinformatics analysis were carried out to screen the 
      differentially expressed lncRNAs and identify the putative target genes 
      separately. LncRNA Malat1 was chosen for further research. Series of loss and 
      gain of functions experiments were performed to study the biological function of 
      LncRNA Malat1. Quantitative real-time PCR (qRT-PCR), Western blot (WB) analysis 
      and immunofluorescence (IF) staining were carried out to separately detect the 
      functions of pancreatic beta-cells at the mRNA and protein levels. Cytoplasmic and 
      nuclear RNA fractionation and fluorescence in situ hybridization (FISH) were used 
      to determine the subcellar location of lncRNA Malat1 in beta-like cells. 
      Enzyme-linked immunosorbent assays (ELISAs) were performed to examine the 
      differentiation and insulin secretion of beta-like cells after stimulation with 
      different glucose concentrations. Structural interactions between lncRNA Malat1 
      and miR-15b-5p and between miR-15b-5p/Ihh were detected by dual luciferase 
      reporter assays (LRAs). RESULTS: We found that the expression of lncRNA Malat1 
      declined during differentiation, and overexpression (OE) of lncRNA Malat1 notably 
      impaired the differentiation and maturation of beta-like cells derived from iPSCs in 
      vitro and in vivo. Most importantly, lncRNA Malat1 could function as a competing 
      endogenous RNA (ceRNA) of miR-15b-5p to regulate the expression of Ihh according 
      to bioinformatics prediction, mechanistic analysis and downstream experiments. 
      CONCLUSION: This study established an unreported regulatory network of lncRNA 
      Malat1 and the miR-15b-5p/Ihh axis during the differentiation of iPSCs into 
      beta-like cells. In addition to acting as an oncogene promoting tumorigenesis, 
      lncRNA Malat1 may be an effective and novel target for treatment of diabetes in 
      the future.
CI  - Copyright (c) 2023. Published by Elsevier Inc.
FAU - Wang, Yao
AU  - Wang Y
AD  - Department of Hepatobiliary and Pancreatic Surgery, Affiliated Hospital of 
      Nantong University, Nantong 226001, China; Research Center of Clinical Medicine, 
      Affiliated Hospital of Nantong University, Nantong 226001, China.
FAU - Ding, Haoxiang
AU  - Ding H
AD  - Nantong University Medical School, Nantong 226001, China.
FAU - Guo, Chengfeng
AU  - Guo C
AD  - Nantong University Medical School, Nantong 226001, China.
FAU - Bao, Qian
AU  - Bao Q
AD  - Nantong University Medical School, Nantong 226001, China.
FAU - Li, Dongqian
AU  - Li D
AD  - Nantong University Medical School, Nantong 226001, China.
FAU - Xiong, Yicheng
AU  - Xiong Y
AD  - Department of Hepatobiliary and Pancreatic Surgery, Affiliated Hospital of 
      Nantong University, Nantong 226001, China. Electronic address: xycdtc@ntu.edu.cn.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20231114
PL  - England
TA  - Cell Signal
JT  - Cellular signalling
JID - 8904683
RN  - 0 (MicroRNAs)
RN  - 0 (RNA, Long Noncoding)
SB  - IM
MH  - *MicroRNAs/genetics/metabolism
MH  - *RNA, Long Noncoding/genetics/metabolism
MH  - *Induced Pluripotent Stem Cells/metabolism
MH  - In Situ Hybridization, Fluorescence
MH  - Cell Differentiation/genetics
OTO - NOTNLM
OT  - Ihh
OT  - Induced pluripotent stem cells (iPSCs)
OT  - lncRNA Malat1
OT  - miR-15b-5p
OT  - beta-like cells
COIS- Declaration of Competing Interest The authors declare that they have no known 
      competing financial interests or personal relationships that could have appeared 
      to influence the work reported in this paper.
EDAT- 2023/11/17 15:26
MHDA- 2023/12/04 12:43
CRDT- 2023/11/16 19:27
PHST- 2023/07/03 00:00 [received]
PHST- 2023/10/18 00:00 [revised]
PHST- 2023/11/13 00:00 [accepted]
PHST- 2023/12/04 12:43 [medline]
PHST- 2023/11/17 15:26 [pubmed]
PHST- 2023/11/16 19:27 [entrez]
AID - S0898-6568(23)00390-X [pii]
AID - 10.1016/j.cellsig.2023.110975 [doi]
PST - ppublish
SO  - Cell Signal. 2024 Jan;113:110975. doi: 10.1016/j.cellsig.2023.110975. Epub 2023 
      Nov 14.