Abstract
A large number of SNPs significant for osteoporosis (OP) had been identified by genome‐wide association studies. However, the underlying association mechanisms were largely unknown. From the perspective of protein phosphorylation, gene expression regulation, and bone cell activity, this study aims to illustrate association mechanisms for representative SNPs of interest.We utilized public databases and bioinformatics tool to identify OP‐associated SNPs which potentially influence protein phosphorylation (phosSNPs). Associations with hip/spine BMD, as well as fracture risk, in human populations for one significant phosSNP, that is, rs227584 (major/minor allele: C/A, EAS population) located in C17orf53 gene, were suggested in prior meta‐analyses. Specifically, carriers of allele C had significant higher BMD and lower risk of low‐trauma fractures than carriers of A. We pursued to test the molecular and cellular functions of rs227584 in bone through osteoblastic cell culture and multiple assays.We identified five phosSNPs significant for OP (P < 0.01). The osteoblastic cells, which was transfected with wild‐type C17orf53 (allele C at rs227584, P126), demonstrated specific interaction with NEK2 kinase, increased expression levels of osteoblastic genes significantly (OPN, OCN, COL1A1, P < 0.05), and promoted osteoblast growth and ALP activity, in contrast to those transfected with mutant C17orf53 (allele A at rs227584, T126).In the light of the consistent evidences between the present functional study in human bone cells and the prior association studies in human populations, we conclude that the SNP rs227584, via altering protein‐kinase interaction, regulates osteoblastic gene expression, influences osteoblast growth and activity, hence to affect BMD and fracture risk in humans.
Highlights
OP is a complex chronic bone disease characterized by reduced bone mass, resulting in microarchitectural deterioration of bone tissue and increased bone fragility.[1]
From the perspective of protein phosphorylation, gene expression, and cellular functions, the present study aims to ascertain the association mechanisms for significant OP‐related SNPs and OP
Based on the bioinformatics prediction results, we carried out the following experiments to validate its impacts on protein molecular functions in osteoblastic cells, including change in protein substrate‐kinase interaction and change in C17orf[53] total protein phosphorylation
Summary
OP is a complex chronic bone disease characterized by reduced bone mass, resulting in microarchitectural deterioration of bone tissue and increased bone fragility.[1]. Plentiful genetic and genomic studies have identified genes that are associated with BMD variation in human populations. From the perspective of protein phosphorylation, gene expression, and cellular functions, the present study aims to ascertain the association mechanisms for significant OP‐related SNPs and OP. Based on the hints from the bioinformatics prediction and previous GWAS studies, we focused our investigation on a representative SNP of interest, that is, phosSNP rs227584, and carried out purposeful downstream assays in vitro to ascertain its molecular and cellular functions. Evidences, collected from the present molecular and cellular study in bone cells, support that SNP rs227584 changes protein phosphorylation site, regulates osteoblastic gene expression, and influences osteoblast functions. The present study highlights the significance of rs227584 and protein phosphorylation in bone biology, and shed new insights into the molecular pathogenesis of OP in humans
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