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. 2022 Oct 28;32(10):1245-1252.
doi: 10.4014/jmb.2208.08042. Epub 2022 Sep 19.

Acceleration of Mesenchymal-to-Epithelial Transition (MET) during Direct Reprogramming Using Natural Compounds

Ji-Hye Seo  1 , Si Won Jang  2 , Young-Joo Jeon  3 , So Young Eun  4 , Yean Ju Hong  5 , Jeong Tae Do  6 , Jung-Il Chae  1 , Hyun Woo Choi  2   7
Affiliations

Acceleration of Mesenchymal-to-Epithelial Transition (MET) during Direct Reprogramming Using Natural Compounds

Ji-Hye Seo et al. J Microbiol Biotechnol. .

Erratum in

Abstract

Induced pluripotent stem cells (iPSCs) can be generated from somatic cells using Oct4, Sox2, Klf4, and c-Myc (OSKM). Small molecules can enhance reprogramming. Licochalcone D (LCD), a flavonoid compound present mainly in the roots of Glycyrrhiza inflata, acts on known signaling pathways involved in transcriptional activity and signal transduction, including the PGC1-α and MAPK families. In this study, we demonstrated that LCD improved reprogramming efficiency. LCD-treated iPSCs (LCD-iPSCs) expressed pluripotency-related genes Oct4, Sox2, Nanog, and Prdm14. Moreover, LCD-iPSCs differentiated into all three germ layers in vitro and formed chimeras. The mesenchymal-to-epithelial transition (MET) is critical for somatic cell reprogramming. We found that the expression levels of mesenchymal genes (Snail2 and Twist) decreased and those of epithelial genes (DSP, Cldn3, Crb3, and Ocln) dramatically increased in OR-MEF (OG2+/+/ROSA26+/+) cells treated with LCD for 3 days, indicating that MET effectively occurred in LCD-treated OR-MEF cells. Thus, LCD enhanced the generation of iPSCs from somatic cells by promoting MET at the early stages of reprogramming.

Keywords: LCD; MET; iPSC; in vitro; natural compound; reprogramming.

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Conflict of interest statement

Conflict of Interest

The authors have no financial conflicts of interest to declare.

Figures

Fig. 1
Fig. 1. LCD enhances reprogramming from fibroblasts.
(A) Oct4-GFP immunofluorescence of reprogrammed MEF with or without LCD. (B) Time-course quantification of GFP+ colonies in reprogrammed MEF at day 7 and day 14 with or without LCD indicated reprogramming efficiency. (C) FACS analysis of Oct4-GFP expression in MEFs after reprogramming at day 14 with or without LCD.
Fig. 2
Fig. 2. Pluripotency of the LCD-iPSCs.
(A) Immunofluorescence staining of core pluripotency markers, Oct4 and Sox2 in LCD-iPSC like ESCs. (B) The qRT-PCR analysis for expression of core pluripotency markers, Oct4, Sox2, Nanog, and Prdm14 in LCD-iPSC like ESCs. (C) The chimeric mouse embryos from LCD-iPSCs. (D) Differentiation potential of LCDiPSCs to three germ layers, stained for markers of ectoderm (Tuji1), mesoderm (α-SMA), and endoderm (Sox17). (E) Genotyping analysis data for detection of differentiation ability of LCD-iPSC in chimeric embryo.
Fig. 3
Fig. 3. LCD promotes MET in early stage of reprogramming.
(A) FACS analysis of Oct4-GFP positive cells in reprogramming cells with LCD (days 1–3, 4–6, 7–9, 10–12, 1–12). (B) Number of Oct4-GFP positive colonies in reprogramming cells with LCD (days 1–3, 4–6, 7–9, 10–12, 1–12). (C) The qRT-PCR analysis for the expression of mesenchymal genes (Snail2, Twist, Zep1, FN1) and (D) epithelial genes (DSP, Cldn3, Crb3, and Ocln).
Fig. 4
Fig. 4. LCD regulates stability of reprogramming factors.
(A) The qRT-PCR analysis of total or endogenous Oct4 and Sox2 expression. (B) Western blot analysis of OCT4 protein expression in LCD-iPSC-like ESCs (day 3, day 6). (C) The pulldown assay demonstrated that LCD can be directly bound to OCT4 and SOX2 proteins.
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References

    1. Takahashi K, Yamanaka SJc. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006;126:663–676. doi: 10.1016/j.cell.2006.07.024. - DOI - PubMed
    1. Thakurela S, Sindhu C, Yurkovsky E, Riemenschneider C, Smith ZD, Nachman I, et al. Differential regulation of OCT4 targets facilitates reacquisition of pluripotency. Nat. Commun. 2019;10:4444. doi: 10.1038/s41467-019-11741-5. - DOI - PMC - PubMed
    1. Hussein SM, Puri MC, Tonge PD, Benevento M, Corso AJ, Clancy JL, et al. Genome-wide characterization of the routes to pluripotency. Nature. 2014;516:198–206. doi: 10.1038/nature14046. - DOI - PubMed
    1. Li R, Liang J, Ni S, Zhou T, Qing X, Li H, et al. A mesenchymal-to-epithelial transition initiates and is required for the nuclear reprogramming of mouse fibroblasts. Cell Stem Cell. 2010;7:51–63. doi: 10.1016/j.stem.2010.04.014. - DOI - PubMed
    1. Zunder ER, Lujan E, Goltsev Y, Wernig M, Nolan GPJCsc. A continuous molecular roadmap to iPSC reprogramming through progression analysis of single-cell mass cytometry. Cell Stem Cell. 2015;16:323–337. doi: 10.1016/j.stem.2015.01.015. - DOI - PMC - PubMed