Long noncoding RNA ANCR inhibits the differentiation of mesenchymal stem cells toward definitive endoderm by facilitating the association of PTBP1 with ID2

Abstract

The generation of definitive endoderm (DE) cells in sufficient numbers is a prerequisite for cell-replacement therapy for liver and pancreatic diseases. Previously, we reported that human adipose-derived mesenchymal stem cells (hAMSCs) can be induced to DE lineages and subsequent functional cells. Clarifying the regulatory mechanisms underlying the fate conversion from hAMSCs to DE is helpful for developing new strategies to improve the differentiation efficiency from hAMSCs to DE organs. Long noncoding RNAs (lncRNAs) have been shown to play pivotal roles in developmental processes, including cell fate determination and differentiation. In this study, we profiled the expression changes of lncRNAs and found that antidifferentiation noncoding RNA ( ANCR ) was downregulated during the differentiation of both hAMSCs and embryonic stem cells (ESCs) to DE cells. ANCR knockdown resulted in the elevated expression of DE markers in hAMSCs, but not in ESCs. ANCR overexpression reduced the efficiency of hAMSCs to differentiate into DE cells. Inhibitor of DNA binding 2 ( ID2 ) was notably downregulated after ANCR knockdown. ID2 knockdown enhanced DE differentiation, whereas overexpression of ID2 impaired this process in hAMSCs. ANCR interacts with RNA-binding polypyrimidine tract-binding protein 1 (PTBP1) to facilitate its association with ID2 mRNA, leading to increased ID2 mRNA stability. Thus, the ANCR /PTBP1/ ID2 network restricts the differentiation of hAMSCs toward DE. Our work highlights the inherent discrepancies between hAMSCs and ESCs. Defining hAMSC-specific signaling pathways might be important for designing optimal differentiation protocols for directing hAMSCs toward DE.

Long noncoding RNA LYPLAL1-AS1 regulates adipogenic differentiation of human mesenchymal stem cells by targeting desmoplakin and inhibiting the Wnt/β-catenin pathway

“…As the majority of LYPLAL1-AS1 was present in the cytoplasm (>70%), we speculate that LYPLAL1-AS1 might be a downstream effector of DSP at the post-transcriptional level. Thus, to evaluate the effect of DSP on LYPLAL1-AS1 mRNA stability, hAMSCs transfected with si-NC or si-DSP were treated with actinomycin D (5 μg/ml), which inhibits RNA polymerase and blocks transcription 17 . Knockdown of DSP improved the RNA stability of LYPLAL1-AS1 (Fig.…”

Crucial roles of different RNA-binding hnRNP proteins in Stem Cells

“…After Pnky and PTBP1 were knocked down respectively, it was found through DEX-Seq 28 sequencing that there were a common part of transcription alterations that affect the differentiation of NSCs and regulate neurogenesis and neuronal development 26 . A new study has shown that PTBP1 can bind and stabilize ID2 mRNA by interacting with lncRNA ANCR, thus inhibiting the differentiation of human adipogenic mesenchymal stem cells (hAMSCs) into defined endoderm cells and reducing the occurrence of liver and pancreatic diseases 29 .…”

Emerging role of lncRNA ELDR in development and cancer

“…Embryonic stem cells differentiate into more specialized cell types to form specific organs by regulating specific gene expression and cell signaling events. Many lncRNAs have been found to regulate signaling molecules in development and differentiation of all the three germ layers to form organs [31]. Those lncRNAs are functionally divergent and perform crucial roles in the generation of cellular diversity and organ complexity during evolution [32].…”