Dynamic expression of microRNAs during the differentiation of human embryonic stem cells into insulin-producing cells

Wei, Rui; Yang, Jin; Liu, Guoqiang; Gao, Mei-juan; Hou, Wenfang; Zhang, Lin; Gao, Hongwei; Liu, Ye; Chen, Guian; Hong, Tianpei

doi:10.1016/j.gene.2013.01.038

Cited by 81 publications

(52 citation statements)

References 54 publications

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“…The results showed that PTGFRN and BMPR2 were the target genes of miR-7 and miR-375, respectively. Thus, the target genes were suitable for screening Wei et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

Identification of target genes for adenohypophysis-prefer miR-7 and miR-375 in cattle

et al. 2015

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ABSTRACT. In this study, expression levels of miRNAs (miRNAs), miR-375 and miR-7, were detected in different tissues of cattle to determine whether adenohypophysis-prefer or exclusively expressed miRNAs, and target genes could be predicted by TargetScan, RNA22, and other software. Target genes related to pituitary function or reproductive traits were identified using a dual-luciferase assay. miR-375 and miR-7 were expressed differently in various tissues. miR-375 and miR-7 showed higher expression in the adenohypophysis, and there was a significant difference compared with expression in other tissues (P < 0.01). The binding sites for miR-7 were the mRNAs of bone morphogenetic protein receptor type II (BMPR2), prostaglandin F2 receptor negative regulator, gonadotropin-releasing hormone receptor, follicle-stimulating hormoneβ, somatostatin receptor 1, and interleukin-1β by bioinformatic analysis; similarly, the mRNAs of BMPR2 and leptin contained binding sites for miR-375, suggesting that these genes are affected by miR-7 or miR-375. Dual-luciferase reporter assays showed that miR-7 regulated (2015) prostaglandin F2 receptor negative regulator expression, while miR-375 regulated BMPR2 expression. The mutated plasmid and miRNA mimics were used to co-transfect NIH3T3 cells; luciferase reporter assays showed that the inhibition of luciferase activity in the wild-type cells dramatically decreased from 75 to 26% with a 3-5-nucleotide mismatch mutation into the seed region of miR-7. miR-375 had nearly lost the ability to inhibit luciferase activity, suggesting that GTCTTCC is the site of interaction between miR-7 and the prostaglandin F2 receptor negative regulator sequence and that GAACAAA is the site of interaction between miR-375 and the BMPR2 sequence.

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“…The results showed that PTGFRN and BMPR2 were the target genes of miR-7 and miR-375, respectively. Thus, the target genes were suitable for screening Wei et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

Identification of target genes for adenohypophysis-prefer miR-7 and miR-375 in cattle

et al. 2015

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“…Wei et al induced hES cells into insulin-producing cells and the dynamic expression of microRNAs during the differentiation was evaluated. They found that miR-375 expression increased from day 4, the maximum expression was detected on day 8, and this then decreased until the end of the differentiation [30]. In the Wei et al study, the maximum expression of miR-375 occurred at the same time that the expression levels of pancreatic hormone encoding genes began to rise [43].…”

Section: Discussionmentioning

confidence: 93%

“…MiR-375 has ten target genes, and some of them are critical for pancreas development, for example Hnf1β, Sox17, GATA6 and Pax6 [30]. .…”

Section: Target Genes Of Mir-375 In the Beta-cell Differentiationmentioning

confidence: 99%

miR-375 induces human decidua basalis-derived stromal cells to become insulin-producing cells

Shaer¹,

Azarpira²,

Vahdati³

et al. 2014

Cellular and Molecular Biology Letters

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This paper focuses on the development of renewable sources of isletreplacement tissue for the treatment of type I diabetes mellitus. Placental tissue-derived mesenchymal stem cells (MSCs) are a promising source for regenerative medicine due to their plasticity and easy availability. They have the potential to differentiate into insulin-producing cells. miR-375 is a micro RNA that is expressed in the pancreas and involved in islet development. Human placental decidua basalis MSCs (PDB-MSCs) were cultured from full-term human placenta. The immunophenotype of the isolated cells was checked for CD90, CD105, CD44, CD133 and CD34 markers. The MSCs (P3) were chemically transfected with hsa-miR-375. Total RNA was extracted 4 and 6 days after transfection. The expressions of insulin, NGN3, GLUT2, PAX4, PAX6, KIR6.2, NKX6.1, PDX1, and glucagon genes were evaluated using real-time qPCR. On day 6, we tested the potency of the clusters in response to the high glucose challenge and assessed the presence of insulin and NGN3 proteins via immunocytochemistry. Flow cytometry analysis confirmed that more than 90% of the cells were positive for CD90, CD105 and CD44 and negative for CD133 and CD34. Morphological changes were followed from day 2. Cell clusters formed during day 6. Insulin-producing clusters showed a deep red color with DTZ. The expression of pancreatic-specific transcription factors increased remarkably during the four days after transfection and significantly increased on day 7. The clusters were positive for insulin and NGN3 proteins, and C-peptide and insulin secretion increased in response to changes in the glucose concentration (2.8 mM and 16.7 mM). In conclusion, the MSCs could be programmed into functional insulin-producing cells by transfection of miR-375.

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“…17 Furthermore, Wei and coworkers induced human ES cells into insulin-producing cells in a stepwise process in vitro and analyzed the expression of some microRNAs during the differentiation process; the pattern was compared with normal development of human pancreatic islets. 21 According to their results, the expression patterns of microRNA-375 and microRNA-7 were similar to the development of human fetal pancreas. Other studies showed that microRNA-7, microRNA-9, microRNA-375, and microRNA-376 had high expression levels during pancreatic islet development in humans.…”

Section: Discussionmentioning

confidence: 94%

“…Human ES cells provide an alternative cell source for regenerative medicine in diabetes because they are characterized by unlimited self-renewal and the ability to differentiate into ILCs. 2,21 Recent studies have shown that the endoderm and pancreatic lineages derived from human ES cells may be a good source for therapeutic purpose. The generation of functional ILCs from human ES and iPS cells with special growth factors also has been investigated.…”

Section: Discussionmentioning

confidence: 99%

Untitled

Shaer

Azarpira

Karimi³

et al. 2016

Exp Clin Transplant

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Objectives: Diabetes results from inadequate insulin production from pancreatic ß-cells. Islet cell replacement is an effective approach for diabetes treatment; however, it is not sufficient for all diabetic patients. Thus, finding a new source with effective maturation of ß-cells is the major goal of many studies. MicroRNAs are a class of small noncoding ribonucleic acid that regulate gene expression through posttranscriptional mechanisms. MicroRNA-7 has high expression level during pancreatic islet development in humans, thereby playing a critical role in pancreatic β-cell function. We study aimed to develop a protocol to differentiate human-induced pluripotent stem cells efficiently into isletlike cell clusters in vitro by using microRNA-7. Materials and Methods: Human-induced pluripotent stem cell colonies were transfected with hsamicroRNA-7 by using siPORT NeoFX transfection agent. Total ribonucleic acid was extracted 24 and 48 hours after transfection. The expression of transcription factors which were important during pancreases development was also performed. On the third day, the potency of the clusters was assessed in response to high glucose levels. Diphenylthiocarbazone was used to identify the existence of the ß-cells. The presence of insulin and Neurogenin-3 proteins was investigated by immunocytochemistry. Results: Morphologic changes were observed on the first day after chemical transfection, and cell clusters were formed on the third day. The expression of pancreatic specific transcription factors was increased on the first day and significantly increased on the second day. The isletlike cell clusters were positive for insulin and Neurogenin-3 proteins in immunocytochemistry. The clusters were stained with Diphenylthiocarbazone and secreted insulin in a glucose challenge test. Conclusions: MicroRNA-7 transcription factor net-work is important in pancreatic endocrine differe-ntiation. Chemical transfection with microRNA-7 can differentiate human induced pluripotent stem cells into functional isletlike cell clusters in a short time.

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Dynamic expression of microRNAs during the differentiation of human embryonic stem cells into insulin-producing cells

Cited by 81 publications

References 54 publications

Identification of target genes for adenohypophysis-prefer miR-7 and miR-375 in cattle

Identification of target genes for adenohypophysis-prefer miR-7 and miR-375 in cattle

miR-375 induces human decidua basalis-derived stromal cells to become insulin-producing cells

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