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Inhibition of the miR-17-92 Cluster Separates Stages of Palatogenesis
Journal article   Open access   Peer reviewed

Inhibition of the miR-17-92 Cluster Separates Stages of Palatogenesis

R J Ries, W Yu, N Holton, H Cao and B A Amendt
Journal of dental research, Vol.96(11), pp.1257-1264
10/2017
DOI: 10.1177/0022034517716915
PMCID: PMC5613877
PMID: 28662367
url
https://doi.org/10.1177/0022034517716915View
Published (Version of record) Open Access

Abstract

The role that noncoding regions of the genome play in the etiology of cleft palate is not well studied. A novel method of microRNA (miR) inhibition that allows for specific miR knockdown in vivo has been developed by our laboratory. To further understand the role of miRs in palatogenesis, we used a new mouse model to inhibit specific miRs within the miR-17-92 cluster. Transgenic mice expressing inhibitory complexes for miR-17 and miR-18 manifested a clefting phenotype that was distinct from that observed in mice carrying inhibitory complexes for miR-17, miR-18, miR-19, and miR-92. An in silico candidate gene analysis and bioinformatics review led us to identify TGFBR2 as a likely target of miR-17 and miR-19 family members. Reverse transcription polymerase chain reaction (RT-PCR) experiments showed that TGFBR1 and TGFBR2 expression levels were elevated in the palates of these miR transgenic embryos at embryonic day 15.5. RT-PCR data also showed that the expression of mature miRs from the miR-17-92 cluster was significantly decreased in the transgenic embryos. Decreased expression of TGFB pathway signaling ligands was also observed. Experiments in cells showed that inhibition of miR-17 and miR-18 was sufficient to induce increases in expression of TGFB receptors, while a concomitant decrease in TGFB signaling ligands was not observed. RT-PCR of mature miR-17-92 in cells demonstrated the selectivity and specificity of inhibitory complexes. While this study builds on previous studies that have implicated miR-17-92 in the regulation of important molecular components of the TGFB signaling pathway, it is likely that interactions remain to be elucidated between miR-17-92 and as-of-yet unidentified molecules important for the control of palatogenesis. The differential regulation of palatogenesis by members of the miR-17-92 cluster indicates that several gene combinations regulate palate elevation and extension during development.
 Phenotype Signal Transduction Receptors, Transforming Growth Factor beta - genetics Animals Protein-Serine-Threonine Kinases - genetics Cleft Palate - embryology Mice, Transgenic Mice MicroRNAs - genetics Reverse Transcriptase Polymerase Chain Reaction Palate - embryology Disease Models, Animal

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