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Identification of New Transcription Factors that Can Promote Pluripotent Reprogramming
Ping Huang1,2,3, Jieying Zhu3,4, Yu Liu5
1The Second School of Clinical Medicine, Southern Medical University, Guangzhou, 510515, Guangdong, China.
Background:
Four 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.
Methods:
The 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.
Results:
ATAC-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.
Conclusions:
In 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.
Insights
New transcription factors TEAD2, TEAD4, and ZIC3 enhance the reprogramming of human somatic cells into induced pluripotent stem cells (iPSCs). These novel iPSCs can differentiate into cardiomyocytes, offering new avenues for regenerative medicine and disease modeling.
Area of Science:
- Stem Cell Biology
- Molecular Biology
- Genetics
Background:
- Induced pluripotent stem cells (iPSCs) are generated using reprogramming factors, with the Yamanaka factors (Oct4, Sox2, Klf4, c-Myc) being the most established.
- Research continues to explore alternative and additional transcription factors to improve reprogramming efficiency and discover novel factor combinations.
- Identifying new factors is crucial for expanding the utility of iPSCs in various applications.
Purpose of the Study:
- To identify novel transcription factors capable of reprogramming human somatic cells, specifically urine-derived cells, into iPSCs.
- To evaluate the efficiency of these novel factors in combination with Yamanaka factors.
- To assess the pluripotency and differentiation potential of the generated iPSCs.
Main Methods:
- Chromatin accessibility (ATAC-seq) and transcriptional profiling (RNA-seq) were performed on human embryonic stem cells and urine cells to identify candidate reprogramming factors.
- Selected transcription factors (TEAD2, TEAD4, ZIC3) were tested for their ability to reprogram urine cells, with reprogramming efficiency quantified.
- Pluripotency of generated iPSCs was confirmed through immunofluorescence, qPCR, RNA-seq, and teratoma formation assays. Differentiation into cardiomyocytes was also assessed in vitro.
Main Results:
- ATAC-seq and RNA-seq data identified TEAD2, TEAD4, and ZIC3 as potential factors involved in urine cell reprogramming.
- Co-expression of TEAD2, TEAD4, and ZIC3 with Yamanaka factors significantly enhanced the reprogramming efficiency of human urine cells.
- The resulting urine-derived iPSCs exhibited pluripotency markers comparable to established human embryonic stem cells and could differentiate into functional cardiomyocytes.
Conclusions:
- TEAD2, TEAD4, and ZIC3 are effective in improving the reprogramming of human urine cells into iPSCs.
- This study provides a new source of stem cells derived from urine for potential clinical applications.
- The newly generated iPSCs are suitable for modeling cardiovascular diseases and developing regenerative therapies.
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