GSE161193
Functional analysis of TMEM53, the disease gene for a novel type of sclerosing bone disorder
Purpose: The goal of this study is to explore the molecular mechanism of the phenotypes found in the TMEM53 mutant human and mouse. Methods: Calvarial mRNA profiles of 3-day-old Tmem53+/- and Tmem53-/- mice were generated by sequencing, in triplicate, using NovaSeq 6000 sequencer. The reads from the RNA-seq were aligned to the mouse genome (NCBI build37.2) using Tophat. Aligned reads were quantified using Cufflinks. Differential gene expression analysis was performed with Cuffdiff. Gene ontology (GO) term enrichment analysis was performed by DAVID 6.8. Results:The gene expression profiling displayed a total of 14,836 genes, including 1,278 differentially expressed genes (DEGs). These DEGs were enriched in some GO terms, and the most enriched term is “ossification”, which is considered to be closely associated with the thickened calvaria in the Tmem53-/- mice. The known marker genes of osteogenic precursors, osteoblasts and osteocytes (Runx2, Sp7, Alpl, Bglap and Ibsp) were up-regulated in  Tmem53-/- mice according to the RNA-seq dataset. The DEGs were also enriched in the reported target gene set of Bmp-Smad pathway. The representative downstream transcriptional targets of Bmp signaling, Id1, Dlx3 and Dlx5, were up-regulated in the calvaria of the Tmem53-/- mice. Conclusions: Our results shows enhanced osteoblastogenesis in the calvaria of the Tmem53-/- mice, which would be attributed to the over-activated Bmp-Smad pathway caused by the Tmem53 deficiency. Overall design: Calvarial mRNA profiles of 3-day-old Tmem53+/- (n=3) and Tmem53-/- mice (n=3)