Alternative splicing in osteoclasts and Paget’s disease of bone

BMC Medical Genetics

Sophie Roux³  Jacques P Brown¹  Laëtitia Michou¹  Stephen McManus³  Martine Bisson³  Gino Laberge³  Roscoe Klinck² 

[1] CHU de Québec, Research Centre and Division of Rheumatology, Department of Medicine, Laval University, Quebec City, PQ, Canada;RNomics platform, Faculty of Medicine, University of Sherbrooke, 3001, 12th Avenue North, Sherbrooke, PQ, Canada;Division of Rheumatology, Department of Medicine, Faculty of Medicine, University of Sherbrooke, 3001, 12th avenue N, Sherbrooke, J1H5N4, PQ, Canada

关键词: p62/SQSTM1;    Paget’s disease of bone;    Osteoclast;    Alternative splicing;   

Others  :  1090984
DOI  :  10.1186/s12881-014-0098-1

 received in 2014-04-11, accepted in 2014-08-07,  发布年份 2014

【 摘 要 】

Background

Mutations in the SQSTM1/p62 gene have been reported in Paget’s disease of bone (PDB), but they are not sufficient to induce the pagetic osteoclast (OC) phenotype. We hypothesized that specific RNA isoforms of OC-related genes may contribute to the overactivity of pagetic OCs, along with other genetic predisposing factors.

Methods

Alternative splicing (AS) events were studied using a PCR-based screening strategy in OC cultures from 29 patients with PDB and 26 healthy donors (HD), all genotyped for the p62^P392L mutation. Primer pairs targeting 5223 characterized AS events were used to analyze relative isoform ratios on pooled cDNA from samples of the four groups (PDB, PDB^P392L, HD, HD^P392L). Of the 1056 active AS events detected in the screening analysis, 192 were re-analyzed on non-amplified cDNA from each subject of the whole cohort.

Results

This analysis led to the identification of six AS events significantly associated with PDB, but none with p62^P392L. The corresponding genes included LGALS8, RHOT1, CASC4, USP4, TBC1D25, and PIDD. In addition, RHOT1 and LGALS8 genes were upregulated in pagetic OCs, as were CASC4 and RHOT1 genes in the presence of p62^P392L. Finally, we showed that the proteins encoded by LGALS8, RHOT1, USP4, TBC1D25, and PIDD were expressed in human OCs.

Conclusion

This study allowed the identification of hitherto unknown players in OC biology, and our findings of a differential AS in pagetic OCs may generate new concepts in the pathogenesis of PDB.

【 授权许可】

   
2014 Klinck et al.; licensee BioMed Central Ltd

【 图 表 】

Figure 1.

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