MicroRNA Profiling During Neural Differentiation of Induced Pluripotent Stem Cells

Abstract

MicroRNAs (miRNA) play an essential role in the regulation of gene expression and influence signaling networks responsible for several cellular processes like differentiation of pluripotent stem cells. Despite several studies on the neurogenesis process, no global analysis of microRNA expression during differentiation of induced pluripotent stem cells (iPSC) to neuronal stem cells (NSC) has been done. Therefore, we compared the profile of microRNA expression in iPSC lines and in NSC lines derived from them, using microarray-based analysis. Two different protocols for NSC formation were used: Direct and two-step via neural rosette formation. We confirmed the new associations of previously described miRNAs in regulation of NSC differentiation from iPSC. We discovered upregulation of miR-10 family, miR-30 family and miR-9 family and downregulation of miR-302 and miR-515 family expression. Moreover, we showed that miR-10 family play a crucial role in the negative regulation of genes expression belonging to signaling pathways involved in neural differentiation: WNT signaling pathway, focal adhesion, and signaling pathways regulating pluripotency of stem cells.

Role of miRNA-mRNA Interaction in Neural Stem Cell Differentiation of Induced Pluripotent Stem Cells

“…As discussed in the results section, the transcriptomic and functional profile of our iPSC-generated NSCs is consistent with the early neuroepithelial cells [ 17 , 27 ]. However the repertoire of 51 DE miRNA between our iPSCs and their differentiated NSCs was smaller than a previous study [ 29 ], likely due to some obvious differences in sample size [ 30 , 31 ], differentiation and miRNA typing methodologies and plausibly due to the heterogeneity of the differentiated NSCs in [ 29 ], as discussed above. miR-9 and miR-124a, which are known to be involved in differentiation of neural stem and progenitor cells into neurons and astrocytes [ 3 , 32 , 33 , 34 , 35 ], were either not expressed or were expressed below the set expression threshold (NRC < 20) in our NSCs.…”

The Significance of MicroRNAs Expression in Regulation of Extracellular Matrix and Other Drug Resistant Genes in Drug Resistant Ovarian Cancer Cell Lines

“…miRNA expression profiling was performed using the microarray approach with Applied BiosystemsTM miRNA 3.1 Array Strip (ThermoFisher Scientific, Waltham, MA, USA). The detailed technical procedure was described earlier [116,117]. Each microarray was designed in accordance with the miRBase Release 17 database, including complementary probes for: 1733 human mature miRNA, 2216 human snoRNA, CDBox RNA, H/ACA Box RNA, and scaRNA, 1658 human pre-miRNA.…”

Extracellular vesicles, microRNA and the preimplantation embryo: non-invasive clues of embryo well-being

“…miRNAs are transcribed ubiquitously across cell types, including embryonic stem cells, and in a cell-or tissue-specific manner (The FANTOM Consortium et al, 2017); in most cases, the top five miRNAs account for about one-half of the total miRNAs (The FANTOM Consortium et al, 2017;O'Brien et al, 2018). The extensive variety of miRNA expressed in a given cell or tissue has empowered comprehensive expression profiling, and has led to the realization that miRNA profiles may be significantly skewed as a result of status changes, including differentiation (Kulcenty et al, 2019) or pathology (Peng and Croce, 2016). Comprehensive miRNA profiling has enabled the prioritization of previously characterized cellular pathways based on their enrichment with proteins prone to transcriptional regulation according to the current expression profile.…”