Prediction and Boolean logical modelling of synergistic microRNA regulatory networks during reprogramming of male germline pluripotent stem cells

Unipotent male germline stem (GS) cells can undergo spontaneous reprogramming to germline pluripotent stem (GPS) cells during in vitro culture. In our previous study, we proposed a Boolean logical model of gene regulatory network (GRN) during reprogramming of GS cells to GPS cells. This study was designed to predict and model synergistic microRNA (miRNA) regulatory network during reprogramming of GS cells into GPS cells. The miRNAs targeting differentially expressed genes (DEGs) among GS and GPS cells were predicted by a novel Gene Ontology (GO) enrichment analysis to construct miRNA synergistic networks (MSN) and identify regulatory miRNA modules. Qualitative Boolean logical model of synergistic miRNAs and its regulated genes was then constructed by considering discrete, asynchronous, multivalued logical formalism using the GINsim modeling and simulation tools. Topology, functional and community overlap studies revealed that mmu-miR-200b-3p, mmu-miR-429-3p and mmu-miR-141-3p, mmu-miR-200a-3p and mmu-miR-200c-3p in MSN belongs to the family of miR-200/429/141 and conjectured to control the pluripotency and reprogramming by promoting Mesenchymal to Epithelial Transition (MET). Synergistic network involving mmu-miR-20b-5p, mmu-miR-20a5p, mmu-miR-106a-5p, mmu-miR-106b-5p, and mmu-miR-17-5p were found to be essential for the maintenance of GS cells. Logical miRNA regulatory network modelling showed that synergistic miRNAs regulates the gene dynamics of MET during GS-GPS reprogramming, as confirmed by perturbation analysis. Taken together, our study predicted novel synergistic miRNAs involved in the regulation of reprogramming and pluripotency in GPS cells. The Boolean logical model of synergistic miRNAs regulatory network further confirms our previous study that gene dynamics of MET regulates GS-GPS reprogramming.

Automated summary

This study predicted and modeled synergistic microRNA regulatory networks, identifying the crucial roles of miR-200/429/141 family in controlling pluripotency and reprogramming, and the essential functions of miR-20/106/17 family in maintaining unipotent cells. The logical modeling demonstrated that synergistic miRNAs regulate MET gene dynamics during reprogramming, which was confirmed by perturbation analysis.