MicroRNA miR-92b-3p regulation of a cardiovascular gene regulatory network in the developing branchial arches

Abstract

ABSTRACTVertebrate branchial arches (BAs) are a developmental paradigm, undergoing coordinated differentiation and morphogenesis to form various adult derivative tissues. MicroRNAs can strengthen gene regulatory networks (GRNs) to promote developmental stability. To interrogate the contribution of microRNAs to BA development, we generated a novel microRNA-sequencing dataset from mouse BAs. We identified 550 expressed microRNAs, of which approximately 20% demonstrate significant differential expression across BA domains. The three most posterior BAs and the connecting outflow tract (PBA/OFT) are enriched in biological processes linked to cardiovascular development. We identified enriched predicted microRNA-target interactions with PBA/OFT upregulated cardiovascular genes and validated transcripts encoding for two fundamental cardiac transcription factors (TFs),Gata6andTbx20,as targets of miR-92b-3p. Furthermore, we demonstrated that miR-92b-3p can downregulate endogenousGATA6andTBX20in human embryonic stem cells (hESCs) undergoing cardiomyocyte differentiation, consistent with conservation of these microRNA-target interactions in a cardiogenic setting. miR-92b-3p has previously been shown to target two other cardiac TFs,Hand2andMef2D.Therefore, we hypothesise that miR-92b-3p acts to stabilise cardiovascular GRNs during PBA/OFT development, through acting in multiple microRNA-mediated coherent feedforward loops.