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Why SNP rs3755955 is associated with human bone mineral density? A molecular and cellular study in bone cells

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Abstract

SNP rs3755955 (major/minor allele: G/A) located in Iduronidase-Alpha-L- (IDUA) gene was reported to be significant for human bone mineral density (BMD). This follow-up study was to uncover the underlying association mechanism through molecular and cellular functional assays relevant to bone. We tested the effects of single nucleotide polymorphisms (SNP) rs3755955 (defined allele G as wild-type and allele A as variant-type) on osteoblastic and osteoclastic functions, as well as protein phosphorylation in stably transfected human fetal osteoblast (hFOB) cell and mononuclear-macrophage (RAW264.7) cell. In hFOB cells, transfection with variant-type IDUA significantly decreased osteoblastic gene expression (OPN, COL1A1 and RANKL) (p < 0.01), impeded cell proliferation (p < 0.05), stimulated cell apoptosis (p < 0.001) and decreased ALP enzyme activity, as compared with that of wild-type IDUA transfection. In RAW264.7 cells, transfection with variant-type IDUA significantly inhibited cell apoptosis (p < 0.01), promoted osteoclastic precursor cell migration (p < 0.0001), growth (p < 0.01), osteoclastic gene expression (TRAP, RANK, Inte-αv and Cath-K) (p < 0.05) and TRAP enzyme activity (p < 0.001), as compared with that of wild-type IDUA transfection. In both hFOB and RAW264.7 cells, the total protein and IDUA protein-specific phosphorylation levels were significantly reduced by variant IDUA transfection, as compared with that of wild-type IDUA transfection (p < 0.05). Variant allele A of phosSNP rs3755955 in IDUA gene regulates protein phosphorylation, inhibits osteoblast function and promotes osteoclastic activity. The SNP rs3755955 could alter IDUA protein phosphorylation, significantly regulates human osteoblastic and osteoclastic gene expression, and influences the growth, differentiation and activity of osteoblast and osteoclast, hence to affect BMD.

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The data used during the current study are available from the corresponding author upon request.

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Acknowledgements

The study was supported by Natural Science Foundation of China (81872681, 31401079 and 81473046), the Science and Technology Project of Suzhou (SS202050, SYS2019024), the QingLan Project of Higher Education of Jiangsu Province and a Project of the Priority Academic Program Development of Jiangsu Higher Education Institutions.

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  1. Pei He and Fei Jiang contributed equally to this work.

Authors and Affiliations

  1. Center for Genetic Epidemiology and Genomics, School of Public Health, Medical College of Soochow University, Suzhou, 215123, Jiangsu, People’s Republic of China

    Pei He, Fei Jiang, Long-Fei Wu, Xu Zhou, Shu-Feng Lei & Fei-Yan Deng

  2. Jiangsu Key Laboratory of Preventive and Translational Medicine for Geriatric Diseases, School of Public Health, Medical College of Soochow University, Suzhou, 215123, Jiangsu, People’s Republic of China

    Pei He, Fei Jiang, Long-Fei Wu, Xu Zhou, Shu-Feng Lei & Fei-Yan Deng

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Contributions

F-YD and S-FL designed and coordinated the study; PH, FJ and L-FW performed the experiments; PH, FJ, and XZ analyzed the data; PH and FJ drafted the manuscript; F-YD revised and finalized the manuscript.

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Correspondence to Fei-Yan Deng.

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11010_2021_4292_MOESM1_ESM.tif

Supplementary file1 (TIF 9760 kb) Supplemental Figure 1. Homologous recombination. A: Vector plasmid used in stable transfection. B: PCR electrophoresis of recombinant plasmids. C: Sequencing results of wild and variant type recombinant plasmids

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He, P., Jiang, F., Wu, LF. et al. Why SNP rs3755955 is associated with human bone mineral density? A molecular and cellular study in bone cells. Mol Cell Biochem 477, 455–468 (2022). https://doi.org/10.1007/s11010-021-04292-1

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