Changes in microRNA expression during differentiation of embryonic and induced pluripotent stem cells to definitive endoderm.
Pluripotent stem cells, including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), have the potential to treat type 1 diabetes through cell replacement therapy. However, the protocols used to generate insulin-expressing cells in vitro frequently result in cells which have an immature phenotype and are functionally restricted. MicroRNAs (miRNAs) are now known to be important in cell fate specification, and a unique miRNA signature characterises pancreatic development at the definitive endoderm stage. Several studies have described differences in miRNA expression between ESCs and iPSCs. Here we have used microarray analysis both to identify miRNAs up- or down-regulated upon endoderm formation, and also miRNAs differentially expressed between ESCs and iPSCs. Several miRNAs fulfilling both these criteria were identified, suggesting that differences in the expression of these miRNAs may affect the ability of pluripotent stem cells to differentiate into definitive endoderm. The expression of these miRNAs was validated by qRT-PCR, and the relationship between one of these miRNAs, miR-151a-5p, and its predicted target gene, SOX17, was investigated by luciferase assay, and suggested an interaction between miR-151a-5p and this key transcription factor. In conclusion, these findings demonstrate a unique miRNA expression pattern for definitive endoderm derived from both embryonic and induced pluripotent stem cells.
Endocrinology Section, Biotherapeutics Department, National Institute of Biological Standards and Control, Blanche Lane, South Mimms, Hertfordshire, EN6 3QG, UK; Section of Cell Biology and Functional Genomics, Division of Diabetes, Endocrinology and Metabolism, Department of Medicine, ICTEM, du Cane Road, Imperial College London, W12 0MN, UK.
Natalie
N
Francis
Endocrinology Section, Biotherapeutics Department, National Institute of Biological Standards and Control, Blanche Lane, South Mimms, Hertfordshire, EN6 3QG, UK.
Melanie
M
Moore
Endocrinology Section, Biotherapeutics Department, National Institute of Biological Standards and Control, Blanche Lane, South Mimms, Hertfordshire, EN6 3QG, UK.
Simona G
SG
Asan
Section of Cell Biology and Functional Genomics, Division of Diabetes, Endocrinology and Metabolism, Department of Medicine, ICTEM, du Cane Road, Imperial College London, W12 0MN, UK.
Guy A
GA
Rutter
Endocrinology Section, Biotherapeutics Department, National Institute of Biological Standards and Control, Blanche Lane, South Mimms, Hertfordshire, EN6 3QG, UK. Electronic address: chris.burns@nibsc.org.
Chris
C
Burns
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D004707:Endoderm / Q000166:cytology / Q000378:metabolism / Q000502:physiology; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D057809:HEK293 Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics*; D046888:Tissue Array Analysis; D014157:Transcription Factors / Q000235:genetics
D035683:MicroRNAs; D014157:Transcription Factors
Definitive endoderm; Differentiation; Embryonic stem cells; Induced pluripotent stem cells; MicroRNA; Pancreas; Type 1 diabetes