{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T05:53:38Z","timestamp":1790661218423,"version":"4.1.0"},"reference-count":35,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,11,8]],"date-time":"2022-11-08T00:00:00Z","timestamp":1667865600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Endocrinol."],"abstract":"<jats:sec><jats:title>Objective<\/jats:title><jats:p>This study aimed to explore the associations of genetic variants in the semaphorin 3A (SEMA3A) signaling pathway genes, including <jats:italic>SEMA3A<\/jats:italic>, <jats:italic>NRP1<\/jats:italic>, <jats:italic>PLXNA1<\/jats:italic>, <jats:italic>PLXNA2<\/jats:italic> and <jats:italic>PLXNA3<\/jats:italic> with osteoporosis (OP) risk and bone mineral density (BMD) in a Chinese Han older adult population.<\/jats:p><\/jats:sec><jats:sec><jats:title>Study design and method<\/jats:title><jats:p>A two-stage design was adopted. Total of 47.8kb regions in the 5 genes were sequenced using targeted next-generation sequencing (NGS) technology in the discovery stage, and the discovered OP-related single nucleotide polymorphisms (SNPs) were further genotyped using improved multiple linkage detection reaction technique in the validation stage. Methods of ALP\/TRAP staining, real-time fluorescent quantitative PCR, and cell proliferation and apoptosis assays were performed with MC3T3-E1 and RAW 264.7 cell lines to clarify biological effects of observed functional variants in cell lines responsible for bone mass remodeling.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Total of 400 postmenopausal women (211 OP cases) were involved in the discovery stage, where 6 common and 4 rare genetic variants were found to be associated with OP risk. In the validation stage among another 859 participants (417 women, 270 OP cases), the <jats:italic>PLXNA2<\/jats:italic> rs2274446 T allele was associated with reduced OP risk and increased femoral neck (FN) BMD compared to the C allele. Moreover, significant associations of <jats:italic>NRP1<\/jats:italic> rs2070296 with FN BMD\/OP risk and of <jats:italic>NRP1<\/jats:italic> rs180868035 with lumbar spine and FN BMDs were also observed in the combination dataset analysis. Compared to the osteoblasts\/osteoclasts transfected with the wild-type <jats:italic>NRP1<\/jats:italic> rs180868035, those transfected with the mutant-type had reduced mRNA expression of osteoblastic genes (i.e., <jats:italic>ALP, RUNX2<\/jats:italic>, <jats:italic>SP7<\/jats:italic> and <jats:italic>OCN<\/jats:italic>), while elevated mRNA expression of osteoclastic genes (i.e., <jats:italic>TRAP<\/jats:italic>, <jats:italic>NFATc1<\/jats:italic> and <jats:italic>CTSK<\/jats:italic>). Furthermore, mutant <jats:italic>NRP1<\/jats:italic> rs180868035 transfection inhibited osteoblast proliferation and osteoclast apoptosis, while promoted osteoclast proliferation and osteoblast apoptosis in corresponding cell lines.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Genetic variants located in <jats:italic>NRP1<\/jats:italic> and <jats:italic>PLXNA2<\/jats:italic> genes were associated with OP risk and BMD. The <jats:italic>NRP1<\/jats:italic> rs180868035 affects bone metabolism by influencing osteoblasts and osteoclasts differentiation, proliferation and apoptosis.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fendo.2022.1014431","type":"journal-article","created":{"date-parts":[[2022,11,8]],"date-time":"2022-11-08T05:00:40Z","timestamp":1667883640000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["Association between SEMA3A signaling pathway genes and BMD\/OP risk: An epidemiological and experimental study"],"prefix":"10.3389","volume":"13","author":[{"given":"Hao-long","family":"Zhou","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mu-hong","family":"Wei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dong-sheng","family":"Di","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ru-yi","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian-li","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ting-ting","family":"Yuan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qian","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ting-ting","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qin","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,11,8]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.2147\/CIA.S54614","article-title":"The epidemiology and management of postmenopausal osteoporosis: a viewpoint from Brazil","volume":"10","author":"Baccaro","year":"2015","journal-title":"Clin Interv Aging"},{"key":"B2","doi-asserted-by":"publisher","first-page":"1","DOI":"10.4158\/GL-2020-0524SUPPL","article-title":"American Association of clinical Endocrinologists\/American college of endocrinology clinical practice guidelines for the diagnosis and treatment of postmenopausal osteoporosis-2020 update","volume":"26","author":"Camacho","year":"2020","journal-title":"Endocr Pract"},{"key":"B3","doi-asserted-by":"publisher","first-page":"609","DOI":"10.1186\/s13018-021-02772-0","article-title":"The global prevalence of osteoporosis in the world: A comprehensive systematic review and meta-analysis","volume":"16","author":"Salari","year":"2021","journal-title":"J Orthop Surg Res"},{"key":"B4","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1007\/s00198-018-4704-5","article-title":"European Guidance for the diagnosis and management of osteoporosis in postmenopausal women","volume":"30","author":"Kanis","year":"2019","journal-title":"Osteoporos Int"},{"key":"B5","doi-asserted-by":"publisher","first-page":"64","DOI":"10.1359\/jbmr.090706","article-title":"Prevalence and trends in low femur bone density among older US adults: NHANES 2005-2006 compared with NHANES III","volume":"25","author":"Looker","year":"2010","journal-title":"J Bone Miner Res"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1002\/jbmr.3757","article-title":"The prevalence of osteoporosis in China, a nationwide, multicenter DXA survey","volume":"34","author":"Zeng","year":"2019","journal-title":"J Bone Miner Res"},{"key":"B7","doi-asserted-by":"publisher","first-page":"5","DOI":"10.4161\/cam.27752","article-title":"Semaphorin 3A: A new player in bone remodeling","volume":"8","author":"Xu","year":"2014","journal-title":"Cell Adh Migr"},{"key":"B8","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1007\/s00395-017-0630-5","article-title":"Sema3A promotes the resolution of cardiac inflammation after myocardial infarction","volume":"112","author":"Rienks","year":"2017","journal-title":"Basic Res Cardiol"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.3390\/cells9092073","article-title":"Osteoblast-osteoclast communication and bone homeostasis","volume":"9","author":"Kim","year":"2020","journal-title":"Cells"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1038\/s41423-021-00637-4","article-title":"SEMA3A-mediated crosstalk between prostate cancer cells and tumor-associated macrophages promotes androgen deprivation therapy resistance","volume":"18","author":"Liu","year":"2021","journal-title":"Cell Mol Immunol"},{"key":"B11","doi-asserted-by":"publisher","first-page":"627","DOI":"10.1016\/j.cmet.2018.12.021","article-title":"Autoregulation of osteocyte Sema3A orchestrates estrogen action and counteracts bone aging","volume":"29","author":"Hayashi","year":"2019","journal-title":"Cell Metab"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1080\/03008207.2021.1974850","article-title":"Sema3A accelerates bone formation during distraction osteogenesis in mice","author":"Zhang","year":"2021","journal-title":"Connect Tissue Res"},{"key":"B13","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1038\/nature11000","article-title":"Osteoprotection by semaphorin 3A","volume":"485","author":"Hayashi","year":"2012","journal-title":"Nature"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1210\/jc.2014-1443","article-title":"Serum Sema3A is in a weak positive association with bone formation marker osteocalcin but not related to bone mineral densities in postmenopausal women","volume":"99","author":"Liu","year":"2014","journal-title":"J Clin Endocrinol Metab"},{"key":"B15","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1016\/S1534-5807(03)00169-2","article-title":"Neuropilin-1 conveys semaphorin and VEGF signaling during neural and cardiovascular development","volume":"5","author":"Gu","year":"2003","journal-title":"Dev Cell"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibs.2008.01.006","article-title":"Semaphorin signaling: progress made and promises ahead","volume":"33","author":"Zhou","year":"2008","journal-title":"Trends Biochem Sci"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1038\/nrd4337","article-title":"Semaphorins and plexins as therapeutic targets","volume":"13","author":"Worzfeld","year":"2014","journal-title":"Nat Rev Drug Discov"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1002\/jbmr.1471","article-title":"PlexinA2 mediates osteoblast differentiation via regulation of Runx2","volume":"27","author":"Oh","year":"2012","journal-title":"J Bone Miner Res"},{"key":"B19","doi-asserted-by":"publisher","first-page":"393","DOI":"10.1002\/dvdy.20512","article-title":"Expression of semaphorin-3A and its receptors in endochondral ossification: potential role in skeletal development and innervation","volume":"234","author":"Gomez","year":"2005","journal-title":"Dev Dyn"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1097\/FPC.0000000000000180","article-title":"A genetic variant in NRP1 is associated with worse response to ranibizumab treatment in neovascular age-related macular degeneration","volume":"26","author":"Lores-Motta","year":"2016","journal-title":"Pharmacogenet Genomics"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gks539","article-title":"SIFT web server: predicting effects of amino acid substitutions on proteins","volume":"40","author":"Sim","year":"2012","journal-title":"Nucleic Acids Res"},{"key":"B22","doi-asserted-by":"publisher","first-page":"144855","DOI":"10.1016\/j.gene.2020.144855","article-title":"Alkaline phosphatase: Structure, expression and its function in bone mineralization","volume":"754","author":"Vimalraj","year":"2020","journal-title":"Gene"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1002\/jcb.22994","article-title":"Signaling networks in RUNX2-dependent bone development","volume":"112","author":"Komori","year":"2011","journal-title":"J Cell Biochem"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.3389\/fcell.2020.601224","article-title":"Recent advances of osterix transcription factor in osteoblast differentiation and bone formation","volume":"8","author":"Liu","year":"2020","journal-title":"Front Cell Dev Biol"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1016\/j.bone.2015.05.046","article-title":"New insights into the biology of osteocalcin","volume":"82","author":"Zoch","year":"2016","journal-title":"Bone"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1002\/jcp.27852","article-title":"Evaluation of efficacy on RANKL induced osteoclast from RAW264.7 cells","volume":"234","author":"Song","year":"2019","journal-title":"J Cell Physiol"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1007\/s00418-018-1636-2","article-title":"Recent advances in osteoclast biology","volume":"149","author":"Ono","year":"2018","journal-title":"Histochem Cell Biol"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1038\/sj.emboj.7601906","article-title":"Structural studies of neuropilin\/antibody complexes provide insights into semaphorin and VEGF binding","volume":"26","author":"Appleton","year":"2007","journal-title":"EMBO J"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.3390\/cells9020517","article-title":"Nrp1 is activated by konjac ceramide binding-induced structural rigidification of the a1a2 domain","volume":"9","author":"Usuki","year":"2020","journal-title":"Cells"},{"key":"B30","doi-asserted-by":"publisher","first-page":"ra31","DOI":"10.1126\/scisignal.2005835","article-title":"DDR2 (discoidin domain receptor 2) suppresses osteoclastogenesis and is a potential therapeutic target in osteoporosis","volume":"8","author":"Zhang","year":"2015","journal-title":"Sci Signal"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1016\/j.yexcr.2005.11.024","article-title":"Neuropilins in neoplasms: expression, regulation, and function","volume":"312","author":"Bielenberg","year":"2006","journal-title":"Exp Cell Res"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1007\/s00198-006-0126-x","article-title":"Association of PLXNA2 polymorphisms with vertebral fracture risk and bone mineral density in postmenopausal Korean population","volume":"17","author":"Hwang","year":"2006","journal-title":"Osteoporos Int"},{"key":"B33","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1016\/j.lfs.2019.01.060","article-title":"Sema6A-plexin-A2 axis stimulates RANKL-induced osteoclastogenesis through PLCgamma-mediated NFATc1 activation","volume":"222","author":"Zhuang","year":"2019","journal-title":"Life Sci"},{"key":"B34","doi-asserted-by":"publisher","first-page":"CD000333","DOI":"10.1002\/14651858.CD000333.pub2","article-title":"Exercise for preventing and treating osteoporosis in postmenopausal women","author":"Howe","year":"2011","journal-title":"Cochrane Database Syst Rev"},{"key":"B35","doi-asserted-by":"publisher","first-page":"458","DOI":"10.1186\/s13018-019-1517-y","article-title":"Associations between serum calcium, 25(OH)D level and bone mineral density in older adults","volume":"14","author":"Liu","year":"2019","journal-title":"J Orthop Surg Res"}],"container-title":["Frontiers in Endocrinology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fendo.2022.1014431\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,11,8]],"date-time":"2022-11-08T05:00:44Z","timestamp":1667883644000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fendo.2022.1014431\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,11,8]]},"references-count":35,"alternative-id":["10.3389\/fendo.2022.1014431"],"URL":"https:\/\/doi.org\/10.3389\/fendo.2022.1014431","relation":{},"ISSN":["1664-2392"],"issn-type":[{"value":"1664-2392","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,11,8]]},"article-number":"1014431"}}