{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,3]],"date-time":"2026-04-03T17:21:35Z","timestamp":1775236895134,"version":"3.50.1"},"reference-count":35,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2021,10,25]],"date-time":"2021-10-25T00:00:00Z","timestamp":1635120000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["PI16\/00991"],"award-info":[{"award-number":["PI16\/00991"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["PI19\/00744"],"award-info":[{"award-number":["PI19\/00744"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["CP15\/00053"],"award-info":[{"award-number":["CP15\/00053"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Semaphorin 4D (Sema4D) is a neurotrophin that is secreted by osteoclasts and binds to its receptor PlexinB1 on osteoblasts to inhibit their differentiation and function. Adenosine A2A activation inhibits osteoclast Sema4D-mediated secretion, diminishes inflammatory osteolysis and prevents bone loss following tenofovir (one of the most used antivirals in HIV). Therefore, tenofovir might activate Sema4D signaling to alter bone turnover. Female C57Bl\/6\/A2AKO mice were ovariectomized and treated with saline (control), tenofovir 75 mg\/Kg\/day, dipyridamole 25 mg\/Kg\/day or a combination for 5 weeks and long bones were prepared for histology. Primary murine-induced osteoclast\/osteoblast were challenged with tenofovir\/dipyridamole 1 \u03bcM each, and the expression of Sema4D\/PlexinB1, RhoA\/ROCK\/IGF1R was studied by RT-PCR, Western blot and immunostaining. In vivo tenofovir showed an increased expression of Sema4D when compared to control mice, and dipyridamole reverted the expression in an A2A-dependent manner. In vitro, tenofovir increases Sema4D expression and secretion in osteoclast precursors, and pre-treatment with dipyridamole reverted this effect. pRhoA and ROCK1 activation were increased and IRS1\/IGF1R expression was diminished by tenofovir in the Vav3\/ARHGAP18 mechanism in osteoblast precursors and reverted by dipyridamole in an A2A-dependent manner. This suggests that tenofovir increases bone loss by activation of Sema4D\/PlexinB1 signaling, which inhibits osteoblast differentiation. Agents that increase local adenosine concentrations, such as dipyridamole, might prevent bone loss following the inhibition of this pathway.<\/jats:p>","DOI":"10.3390\/ijms222111490","type":"journal-article","created":{"date-parts":[[2021,10,25]],"date-time":"2021-10-25T21:40:21Z","timestamp":1635198021000},"page":"11490","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Tenofovir Modulates Semaphorin 4D Signaling and Regulates Bone Homeostasis, Which Can Be Counteracted by Dipyridamole and Adenosine A2A Receptor"],"prefix":"10.3390","volume":"22","author":[{"given":"Patricia","family":"Llamas-Granda","sequence":"first","affiliation":[{"name":"Bone and Joint Research Unit, IIS-Fundaci\u00f3n Jim\u00e9nez D\u00edaz UAM, 28040 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laura","family":"Martin-Rodr\u00edguez","sequence":"additional","affiliation":[{"name":"Bone and Joint Research Unit, IIS-Fundaci\u00f3n Jim\u00e9nez D\u00edaz UAM, 28040 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6525-2944","authenticated-orcid":false,"given":"Raquel","family":"Largo","sequence":"additional","affiliation":[{"name":"Bone and Joint Research Unit, IIS-Fundaci\u00f3n Jim\u00e9nez D\u00edaz UAM, 28040 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3241-991X","authenticated-orcid":false,"given":"Gabriel","family":"Herrero-Beaumont","sequence":"additional","affiliation":[{"name":"Bone and Joint Research Unit, IIS-Fundaci\u00f3n Jim\u00e9nez D\u00edaz UAM, 28040 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5368-574X","authenticated-orcid":false,"given":"Ar\u00e1nzazu","family":"Mediero","sequence":"additional","affiliation":[{"name":"Bone and Joint Research Unit, IIS-Fundaci\u00f3n Jim\u00e9nez D\u00edaz UAM, 28040 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,10,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"263","DOI":"10.7150\/ijms.5.263","article-title":"Eph receptors and ephrin signaling pathways: A role in bone homeostasis","volume":"5","author":"Edwards","year":"2008","journal-title":"Int. J. Med. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"837","DOI":"10.1002\/jbmr.2421","article-title":"Netrin-1 is a critical autocrine\/paracrine factor for osteoclast differentiation","volume":"30","author":"Mediero","year":"2015","journal-title":"J. Bone Min. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1016\/S0006-8993(00)02700-1","article-title":"Expression of mRNA for axon guidance molecules, such as semaphorin-III, netrins and neurotrophins, in human osteoblasts and osteoclasts","volume":"878","author":"Togari","year":"2000","journal-title":"Brain Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1473","DOI":"10.1038\/nm.2489","article-title":"Suppression of bone formation by osteoclastic expression of semaphorin 4D","volume":"17","author":"Shinohara","year":"2011","journal-title":"Nat. Med."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Dacquin, R., Domenget, C., Kumanogoh, A., Kikutani, H., Jurdic, P., and Machuca-Gayet, I. (2011). Control of bone resorption by semaphorin 4D is dependent on ovarian function. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0026627"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"860","DOI":"10.1177\/0022034513500306","article-title":"Advances in the regulation of osteoclasts and osteoclast functions","volume":"92","author":"Boyce","year":"2013","journal-title":"J. Dent. Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/S0065-2776(06)93003-X","article-title":"Immune semaphorins: Increasing members and their diverse roles","volume":"93","author":"Kikutani","year":"2007","journal-title":"Adv. Immunol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.clim.2015.12.012","article-title":"Soluble SEMA4D\/CD100: A novel immunoregulator in infectious and inflammatory diseases","volume":"163","author":"Maleki","year":"2016","journal-title":"Clin. Immunol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1186\/1476-4598-9-251","article-title":"Roles of Sema4D and Plexin-B1 in tumor progression","volume":"9","author":"Kumanogoh","year":"2010","journal-title":"Mol. Cancer"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1093\/intimm\/dxh224","article-title":"Soluble CD100 functions on human monocytes and immature dendritic cells require plexin C1 and plexin B1, respectively","volume":"17","author":"Pasterkamp","year":"2005","journal-title":"Int. Immunol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"5262","DOI":"10.4049\/jimmunol.157.12.5262","article-title":"CD100 is associated with CD45 at the surface of human T lymphocytes. Role in T cell homotypic adhesion","volume":"157","author":"Herold","year":"1996","journal-title":"J. Immunol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3463","DOI":"10.1242\/jcs.00674","article-title":"Immune semaphorins: A new area of semaphorin research","volume":"116","author":"Kumanogoh","year":"2003","journal-title":"J. Cell Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1027","DOI":"10.1093\/intimm\/dxg098","article-title":"Involvement of CD100, a lymphocyte semaphorin, in the activation of the human immune system via CD72: Implications for the regulation of immune and inflammatory responses","volume":"15","author":"Ishida","year":"2003","journal-title":"Int. Immunol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3498","DOI":"10.1182\/blood.V97.11.3498","article-title":"Functional soluble CD100\/sema4D released from activated lymphocytes: Possible role in normal and pathologic immune responses","volume":"97","author":"Wang","year":"2001","journal-title":"Blood"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1481","DOI":"10.1002\/art.39086","article-title":"Semaphorin 4D Contributes to Rheumatoid Arthritis by Inducing Inflammatory Cytokine Production: Pathogenic and Therapeutic Implications","volume":"67","author":"Yoshida","year":"2015","journal-title":"Arthritis Rheumatol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"745","DOI":"10.1182\/blood-2010-12-324848","article-title":"Expansion of CD8+ T cells lacking Sema4D\/CD100 during HIV-1 infection identifies a subset of T cells with decreased functional capacity","volume":"119","author":"Eriksson","year":"2012","journal-title":"Blood"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Correa-Rocha, R., Lopez-Abente, J., Gutierrez, C., Perez-Fernandez, V.A., Prieto-Sanchez, A., Moreno-Guillen, S., Mu\u00f1oz-Fernandez, M.A., and Pion, M. (2018). CD72\/CD100 and PD-1\/PD-L1 markers are increased on T and B cells in HIV-1+ viremic individuals, and CD72\/CD100 axis is correlated with T-cell exhaustion. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0203419"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1016\/j.clim.2017.09.010","article-title":"Low levels of the immunoregulator Semaphorin 4D (CD100) in sera of HIV patients","volume":"191","author":"Vadasz","year":"2018","journal-title":"Clin. Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1063","DOI":"10.3851\/IMP1874","article-title":"Bone turnover, osteoprotegerin\/RANKL and inflammation with antiretroviral initiation: Tenofovir versus non-tenofovir regimens","volume":"16","author":"Brown","year":"2011","journal-title":"Antivir. Ther."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Feig, J.L., Mediero, A., Corciulo, C., Liu, H., Zhang, J., Perez-Aso, M., Picard, L., Wilder, T., and Cronstein, B. (2017). The antiviral drug tenofovir, an inhibitor of Pannexin-1-mediated ATP release, prevents liver and skin fibrosis by downregulating adenosine levels in the liver and skin. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0188135"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"96","DOI":"10.3389\/fphys.2014.00096","article-title":"Role of Pannexin-1 hemichannels and purinergic receptors in the pathogenesis of human diseases","volume":"5","author":"Velasquez","year":"2014","journal-title":"Front. Physiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"923","DOI":"10.1002\/jbmr.3665","article-title":"Tenofovir Causes Bone Loss via Decreased Bone Formation and Increased Bone Resorption, Which Can Be Counteracted by Dipyridamole in Mice","volume":"34","author":"Wilder","year":"2019","journal-title":"J. Bone Min. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1002\/jbm.b.33561","article-title":"Bone regeneration in critical bone defects using three-dimensionally printed beta-tricalcium phosphate\/hydroxyapatite scaffolds is enhanced by coating scaffolds with either dipyridamole or BMP-2","volume":"105","author":"Ishack","year":"2015","journal-title":"J. Biomed. Mater. Res. B Appl. Biomater."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3487","DOI":"10.1096\/fj.201700217R","article-title":"Adenosine A2A receptor (A2AR) stimulation modulates expression of semaphorins 4D and 3A, regulators of bone homeostasis","volume":"32","author":"Mediero","year":"2018","journal-title":"FASEB J."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.semcdb.2012.09.008","article-title":"Semaphorins in bone development, homeostasis, and disease","volume":"24","author":"Kang","year":"2013","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2165","DOI":"10.1097\/QAD.0b013e32801022eb","article-title":"Antiretroviral therapy and the prevalence of osteopenia and osteoporosis: A meta-analytic review","volume":"20","author":"Brown","year":"2006","journal-title":"AIDS"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1097\/QAI.0b013e318289bb7e","article-title":"Functional impairment is associated with low bone and muscle mass among persons aging with HIV infection","volume":"63","author":"Erlandson","year":"2013","journal-title":"J. Acquir. Immune Defic. Syndr."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1097\/QAI.0000000000002207","article-title":"Management of Osteoporosis in Patients Living With HIV-A Systematic Review and Meta-analysis","volume":"83","author":"Rosendahl","year":"2020","journal-title":"J. Acquir. Immune Defic. Syndr."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1007\/s10067-018-4089-z","article-title":"Rheumatic diseases in HIV-infected patients in the post-antiretroviral therapy era: A tertiary care center experience","volume":"38","author":"Parperis","year":"2019","journal-title":"Clin. Rheumatol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1038\/s41413-018-0019-6","article-title":"Paracrine and endocrine actions of bone\u2014the functions of secretory proteins from osteoblasts, osteocytes, and osteoclasts","volume":"6","author":"Han","year":"2018","journal-title":"Bone Res."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Callebaut, C., Liu, Y., Babusis, D., Ray, A., Miller, M., and Kitrinos, K. (2017). Viability of primary osteoblasts after treatment with tenofovir alafenamide: Lack of cytotoxicity at clinically relevant drug concentrations. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0169948"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.bbrc.2010.02.080","article-title":"Tenofovir treatment of primary osteoblasts alters gene expression profiles: Implications for bone mineral density loss","volume":"394","author":"Grigsby","year":"2010","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1007\/s10517-017-3825-8","article-title":"Involvement of Semaphorin (Sema4D) in T-Dependent Activation of B Cells","volume":"163","author":"Kuklina","year":"2017","journal-title":"Bull. Exp. Biol. Med."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Kruppke, B., Ray, S., Alt, V., Rohnke, M., Kern, C., Kampschulte, M., Heinemann, C., Budak, M., Adam, J., and D\u00f6hner, N. (2020). Gelatin-Modified Calcium\/Strontium Hydrogen Phosphates Stimulate Bone Regeneration in Osteoblast\/Osteoclast Co-Culture and in Osteoporotic Rat Femur Defects-In Vitro to In Vivo Translation. Molecules, 25.","DOI":"10.3390\/molecules25215103"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e45","DOI":"10.1093\/nar\/29.9.e45","article-title":"A new mathematical model for relative quantification in real-time RT-PCR","volume":"29","author":"Pfaffl","year":"2001","journal-title":"Nucleic Acids Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/21\/11490\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:23:01Z","timestamp":1760167381000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/21\/11490"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,10,25]]},"references-count":35,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2021,11]]}},"alternative-id":["ijms222111490"],"URL":"https:\/\/doi.org\/10.3390\/ijms222111490","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,10,25]]}}}