Computational and functional characterization of four SNPs in the SOST locus associated with osteoporosis

Ye, Weiyuan; Wang, Ya; Mei, Bing; Hou, Sasa; Liu, Xinhong; Wu, Guiju; Qin, Longjuan; Zhao, Kehui; Huang, Qingyang

doi:10.1016/j.bone.2018.01.001

Cited by 18 publications

(21 citation statements)

References 69 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In particular, miR-98 binds to SOST mRNA. leading to osteoblast differentiation (24). miR-335 and miR-433 expression bind to dkk1 mRNA in its 3'UTR, thus promoting osteoblastogenesis (25,26).…”

Section: Discussionmentioning

confidence: 99%

miR‑483‑3p promotes the osteogenesis of human osteoblasts by�targeting Dikkopf 2 (DKK2) and the Wnt signaling pathway

et al. 2020

View full text Add to dashboard Cite

Osteoporosis is a systemic metabolic bone disease during which bone mass decreases and bone quality is reduced. Maintaining the bone formation capacity of osteoblasts is crucial for the treatment of osteoporosis. In the present study, bioinformatics analysis was performed on online microarray expression profiles to identify miRNA(s) related to osteoblast proliferation and bone marrow-derived mesenchymal stem cell (BMSc) osteogenic differentiation. The specific effects of candidate miRNAs on cell proliferation, osteogenic differentiation and Wnt signaling-related factors were examined. As regards the downstream mechanisms, online tools were employed to predict the downstream targets of candidate miRNAs and the predicted miRNA-mRNA binding was verified. Finally, the dynamic effects of miRNAs and mRNAs were examined. The results revealed that miR-483-3p expression was decreased in bone tissue samples from patients with osteoporosis. In miR-483-3p-overexpressing human osteoblasts, cell viability, dNA synthesis capacity and osteogenesis were promoted, and the protein levels of Wnt1, β-catenin and cyclin d1 were increased. However, the protein receptor activator of nuclear factor kappa-Β ligand (RANKL)/osteoprotegerin (OPG) ratio and cell apoptotic rate were decreased. The Wnt signaling, antagonist dikkopf 2 (dKK2), was targeted and negatively regulated by miR-483-3p. dKK2 knockdown exerted similar effects as miR-483-3p overexpression, while dKK2 overexpression inhibited cell viability, dNA synthesis capacity and osteogenesis. dKK2 overexpression also decreased the Wnt1, β-catenin, and cyclin d1 protein levels, whereas it promoted the the RANKL/OPG ratio and the apoptosis of human osteoblasts. dKK2 overexpression reversed the functions of miR-483-3p overexpression. On the whole, the findings of the present study demonstrate that the miR-483-3p/dKK2 axis modulates the bone formation process by affecting osteoblast proliferation, pre-osteoblast differentiation into mature osteoblasts and new bone matrix formation.

show abstract

Section: Discussionmentioning

confidence: 99%

miR‑483‑3p promotes the osteogenesis of human osteoblasts by�targeting Dikkopf 2 (DKK2) and the Wnt signaling pathway

et al. 2020

View full text Add to dashboard Cite

show abstract

“…A human osteoblast cell line hFOB1.19, two human osteosarcoma cell lines (Saos‐2 and U‐2OS), and a human embryonic kidney cell line HEK293T were cultured as described previously (Ye et al, ). Cell identities were verified.…”

Section: Methodsmentioning

confidence: 99%

“…Collection, lysis, protein concentration determination and immunoblotting of cultured cells were performed as described previously (Ye et al, ). The primary antibodies were purchased from the following sources: Anti‐USF3 (HPA029245, Atlas Antibodies; sc‐99481; Santa Cruz), anti‐RUNX3 (A7303; ABclonal), anti‐SMAD1 (10429‐1‐AP; Proteintech), and anti‐TUBULIN (sc‐5546; Santa Cruz).…”

Section: Methodsmentioning

confidence: 99%

Osteoporosis genome‐wide association study variant c.3781 C>A is regulated by a novel anti‐osteogenic factor miR‐345‐5p

Liu

et al. 2020

Human Mutation

Self Cite

View full text Add to dashboard Cite

Upstream transcription factor family member 3 (USF3) c.3781C>A (rs1026364) in the 3′-untranslated region (3′-UTR) has been firmly associated with bone mineral density (BMD) in genome-wide association study (GWAS). However, the molecular mechanism by which it influences BMD and osteoporosis is unknown. Bioinformatics analyses suggested that the risk c.3781A allele creates a target site for hsa-miR-345-5pbinding. Luciferase assay validated that the c.3781A allele displayed significantly lower luciferase activities than the c.3781C allele in the human osteoblast cell line hFOB1.19, osteosarcoma cell lines U-2OS and Saos-2, and embryonic kidney cell line 293T. Furthermore, hsa-miR-345-5p regulated USF3 expression on both messenger RNA and protein levels in hFOB1.19 and U937 cells with heterozygous A/C genotype.Transfection of hsa-miR-345-5p antagomiR in heterozygous hFOB1.19 cells significantly increased the expression of osteogenic marker genes RUNX2, OSTERIX, COL1A1, ALP, OPN, OCN, and alkaline phosphatase activity and matrix mineralization level. Importantly, we found that hsa-miR-345-5p also inhibits osteoblast maturation in cell lines U-2OS with hsa-miR-345-5p nonbinding C/C genotype by targeting RUNX3 and SMAD1. Our findings uncovered a novel pathogenetic mechanism of osteoporosis by GWAS variant c.3781C>A-mediated disruption of hsa-miR-345-5p binding at the 3′-UTR of USF3 and the functional role of hsa-miR-345-5p in osteogenic differentiation. K E Y W O R D SGWAS SNP, miR-345-5p, osteoporosis, RUNX3, SMAD1, USF3

show abstract

“…7 Recent functional evidence has also shown that SNPs in this area (one of which, rs7220711, is in high LD [r 2 =0·99] with rs7209826) regulate SOST expression via differential transcription factor binding. 28 Previous Mendelian randomisation studies have also made use of non-coding variants with an effect on gene expression as proxies for pharmacologic modulation of the same target-gene. 22,26,29,30 We extracted estimates for these two SNPs (rs7209826 and rs188810925) from GWAS consortia listed in Table S2 in the appendix.…”

Section: Genetic Instrument Selection and Validationmentioning

confidence: 99%

Lifelong genetically lowered sclerostin and risk of cardiovascular disease

Bovijn

Krebs

Chen

et al. 2019

Preprint

View full text Add to dashboard Cite

BackgroundInhibition of sclerostin is a novel therapeutic approach to lowering fracture risk. However, phase III randomised controlled trials (RCTs) of romosozumab, a monoclonal antibody that inhibits sclerostin, suggest an imbalance of serious cardiovascular events. MethodsWe used two independent genetic variants (rs7209826 and rs188810925) in SOST (encoding sclerostin) associated with bone mineral density (BMD) as proxies for therapeutic inhibition of sclerostin. We estimated the effects on risk of osteoporosis, fracture, coronary heart disease (CHD) and a further 22 cardiometabolic risk factors and diseases, by combining data from up to 478,967 participants of European ancestry from three prospective cohorts and up to 1,030,836 participants from nine GWAS consortia. In addition, we performed meta-analyses of cardiovascular outcome data from phase III RCTs of romosozumab. ResultsMeta-analysis of RCTs identified a higher risk of cardiac ischemic events in patients randomised to romosozumab (25 events among 4,298 individuals; odds ratio [OR] 2·98; 95% confidence interval [CI], 1·18 to 7·55; P=0·017). Scaled to the equivalent dose of romosozumab (210mg/month; 0·09 g/cm 2 higher BMD), the SOST variants associated with lower risk of fracture (OR, 0·59; 95% CI, 0·54-0·66; P= 1·4×10 -24 ), and osteoporosis (OR, 0·43; 95% CI, 0·36-0·52; P=2·4×10 -18 ). The SOST variants associated with higher risk of myocardial infarction and/or coronary revascularisation (69,649 cases; OR, 1·18; 95% CI, 1·06-1·32; P=0·003) and type 2 diabetes (OR 1·15; 95% CI, 1·05-1·27; P=0·003), higher systolic blood pressure (1·3mmHg; 95% CI 0·8-1·9; P=5·9×10 -6 ) and waist-to-hip-ratio adjusted for BMI (0·05 SDs; 95% CI, 0·02 to 0·08; P=8·5×10 -4 ). ConclusionsGenetically and therapeutically lowered sclerostin leads to higher risk of cardiovascular events. Rigorous evaluation of the cardiovascular safety of romosozumab and other sclerostin inhibitors is warranted.

show abstract

Computational and functional characterization of four SNPs in the SOST locus associated with osteoporosis

Cited by 18 publications

References 69 publications

miR‑483‑3p promotes the osteogenesis of human osteoblasts by�targeting Dikkopf 2 (DKK2) and the Wnt signaling pathway

miR‑483‑3p promotes the osteogenesis of human osteoblasts by�targeting Dikkopf 2 (DKK2) and the Wnt signaling pathway

Osteoporosis genome‐wide association study variant c.3781 C>A is regulated by a novel anti‐osteogenic factor miR‐345‐5p

Lifelong genetically lowered sclerostin and risk of cardiovascular disease

Contact Info

Product

Resources

About