Abstract
BackgroundFour transcription factors, Oct4, Sox2, Klf4, and c-Myc (the Yamanka factors), can reprogram somatic cells to induced pluripotent stem cells (iPSCs). Many studies have provided a number of alternative combinations to the non-Yamanaka factors. However, it is clear that many additional transcription factors that can generate iPSCs remain to be discovered.MethodsThe chromatin accessibility and transcriptional level of human embryonic stem cells and human urine cells were compared by Assay for Transposase-Accessible Chromatin with high-throughput sequencing (ATAC-seq) and RNA sequencing (RNA-seq) to identify potential reprogramming factors. Selected transcription factors were employed to reprogram urine cells, and the reprogramming efficiency was measured. Urine-derived iPSCs were detected for pluripotency by Immunofluorescence, quantitative polymerase chain reaction, RNA sequencing and teratoma formation test. Finally, we assessed the differentiation potential of the new iPSCs to cardiomyocytes in vitro.ResultsATAC-seq and RNA-seq datasets predicted TEAD2, TEAD4 and ZIC3 as potential factors involved in urine cell reprogramming. Transfection of TEAD2, TEAD4 and ZIC3 (in the presence of Yamanaka factors) significantly improved the reprogramming efficiency of urine cells. We confirmed that the newly generated iPSCs possessed pluripotency characteristics similar to normal H1 embryonic stem cells. We also confirmed that the new iPSCs could differentiate to functional cardiomyocytes.ConclusionsIn conclusion, TEAD2, TEAD4 and ZIC3 can increase the efficiency of reprogramming human urine cells into iPSCs, and provides a new stem cell sources for the clinical application and modeling of cardiovascular disease.Graphical abstract
Highlights
Four transcription factors, Oct4, Sox2, Klf4, and c-Myc, can reprogram somatic cells to induced pluripotent stem cells
We detected binding motifs for ZIC3, which has previously been demonstrated as a key regulator in maintain mouse embryonic stem cells, and could enhance generate mouse induced pluripotent stem cells [29, 30]
When looked at the gene expression data, TEAD2, TEAD4 and ZIC3 were higher expressed in H1, the expression of TEAD1 and TEAD3 in urine cells (UCs) were similar with H1 (Fig. 1d)
Summary
Oct, Sox, Klf, and c-Myc (the Yamanka factors), can reprogram somatic cells to induced pluripotent stem cells (iPSCs). The induced pluripotent stem cell (iPSC) technology pioneered by Yamanaka et al [1] was one of the most exciting discoveries in biotechnology this century. This technology has great potential in precision medicine, disease modeling, and drug discovery, and it avoids ethical issues related to the generation of human embryonic stem cells (hESCs) from human embryos. The classic Yamanaka factors used to reprogram include the four transcription factors (TFs): OCT4, SOX2, KLF4, and c-MYC (OSKM) [1, 2]. Yu et al [3] used the OCT4, SOX2, NANOG and LIN28 systems to generate human iPSCs, and Wang et al [4] demonstrated the combination of Jdp, Jhdm1b, Mkk, Glis, Nanog, Essrb, and Sall (7F) for mouse iPSCs reprogramming
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