{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T12:15:29Z","timestamp":1792498529194,"version":"4.5.0"},"reference-count":52,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2021,3,23]],"date-time":"2021-03-23T00:00:00Z","timestamp":1616457600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003399","name":"Science and Technology Commission of Shanghai Municipality","doi-asserted-by":"publisher","award":["18PJD056"],"award-info":[{"award-number":["18PJD056"]}],"id":[{"id":"10.13039\/501100003399","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003399","name":"Science and Technology Commission of Shanghai Municipality","doi-asserted-by":"publisher","award":["16PJ1405500"],"award-info":[{"award-number":["16PJ1405500"]}],"id":[{"id":"10.13039\/501100003399","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Multicentric carpotarsal osteolysis (MCTO) is a rare skeletal dysplasia with osteolysis at the carpal and tarsal bones. Heterozygous missense mutations in the transcription factor MAFB are found in patients with MCTO. MAFB is reported to negatively regulate osteoclastogenesis in vitro. However, the in vivo function of MAFB and its relation to MCTO remains unknown. In this study, we generated zebrafish MAFB homolog mafbb mutant utilizing CRISPR\/Cas9 technology. Mafbb deficient zebrafish demonstrated enhanced osteoclast cell differentiation and abnormal cartilage and bone development resembling MCTO patients. It is known that osteoclasts are hematopoietic cells derived from macrophages. Loss of mafbb caused selective expansion of definitive macrophages and myeloid cells, supporting that mafbb restricts myeloid differentiation in vivo. We also demonstrate that MAFB MCTO mutations failed to rescue the defective osteoclastogenesis in mafbb\u2212\/\u2212 embryos, but did not affect osteoclast cells in wild type embryos. The mechanism of MCTO mutations is likely haploinsufficiency. Zebrafish mafbb mutant provides a useful model to study the function of MAFB in osteoclastogenesis and the related MCTO disease.<\/jats:p>","DOI":"10.3390\/biom11030480","type":"journal-article","created":{"date-parts":[[2021,3,23]],"date-time":"2021-03-23T14:08:16Z","timestamp":1616508496000},"page":"480","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Zebrafish mafbb Mutants Display Osteoclast Over-Activation and Bone Deformity Resembling Osteolysis in MCTO Patients"],"prefix":"10.3390","volume":"11","author":[{"given":"Yujie","family":"Han","sequence":"first","affiliation":[{"name":"Engineering Research Center of Cell &amp; Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weihao","family":"Shao","sequence":"additional","affiliation":[{"name":"Engineering Research Center of Cell &amp; Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dan","family":"Zhong","sequence":"additional","affiliation":[{"name":"Engineering Research Center of Cell &amp; Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cui","family":"Ma","sequence":"additional","affiliation":[{"name":"Engineering Research Center of Cell &amp; Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaona","family":"Wei","sequence":"additional","affiliation":[{"name":"Engineering Research Center of Cell &amp; Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abrar","family":"Ahmed","sequence":"additional","affiliation":[{"name":"Engineering Research Center of Cell &amp; Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tingting","family":"Yu","sequence":"additional","affiliation":[{"name":"Shanghai Children\u2019s Medical Center, Department of Medical Genetics and Molecular Diagnostic Laboratory, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Jing","sequence":"additional","affiliation":[{"name":"Department of Hepatobiliary Pancreatic Surgery, Shanghai Changhai Hospital, Shanghai 200433, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7726-511X","authenticated-orcid":false,"given":"Lili","family":"Jing","sequence":"additional","affiliation":[{"name":"Engineering Research Center of Cell &amp; Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,3,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1538\/expanim.19-0076","article-title":"Role of MafB in macrophages","volume":"69","author":"Hamada","year":"2020","journal-title":"Exp. Anim."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1848","DOI":"10.1016\/j.jid.2016.05.088","article-title":"Transcription Factor MafB Coordinates Epidermal Keratinocyte Differentiation","volume":"136","author":"Miyai","year":"2016","journal-title":"J. Investig. Dermatol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1093\/jb\/mvg130","article-title":"The mouse mafB 5\u2032-upstream fragment directs gene expression in myelomonocytic cells, differentiated macrophages and the ventral spinal cord in transgenic mice","volume":"134","author":"Michito","year":"2003","journal-title":"J. Biochem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1987","DOI":"10.1093\/emboj\/19.9.1987","article-title":"MafB is an inducer of monocytic differentiation","volume":"19","author":"Kelly","year":"2000","journal-title":"EMBO J."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1700","DOI":"10.1038\/s41467-017-01711-0","article-title":"MafB is a critical regulator of complement component C1q","volume":"8","author":"Tran","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5715","DOI":"10.1128\/MCB.00001-06","article-title":"MafB is essential for renal development and F4\/80 expression in macrophages","volume":"26","author":"Moriguchi","year":"2006","journal-title":"Mol. Cell Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3253","DOI":"10.1182\/blood-2006-09-048249","article-title":"MafB negatively regulates RANKL-mediated osteoclast differentiation","volume":"109","author":"Kim","year":"2007","journal-title":"Blood"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1038\/nature01658","article-title":"Osteoclast differentiation and activation","volume":"423","author":"Boyle","year":"2003","journal-title":"Nature"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"626","DOI":"10.2174\/1574888X10666151019115724","article-title":"Osteoclasts and Remodeling Based Bone Formation","volume":"11","author":"Kylmaoja","year":"2016","journal-title":"Curr. Stem Cell Res. Ther."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/j.bbrc.2016.05.019","article-title":"Osteoclast function and bone-resorbing activity: An overview","volume":"476","author":"Soysa","year":"2016","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"11","DOI":"10.4248\/BR201301003","article-title":"Osteoclasts: New Insights","volume":"1","author":"Xu","year":"2013","journal-title":"Bone Res."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Anesi, A., Generali, L., Sandoni, L., Pozzi, S., and Grande, A. (2019). From Osteoclast Differentiation to Osteonecrosis of the Jaw: Molecular and Clinical Insights. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20194925"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/S0092-8674(00)81569-X","article-title":"Osteoprotegerin Ligand Is a Cytokine that Regulates Osteoclast Differentiation and Activation","volume":"93","author":"Lacey","year":"1998","journal-title":"Cell"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2287","DOI":"10.1002\/ajmg.a.36641","article-title":"Multicentric carpotarsal osteolysis syndrome is caused by only a few domain-specific mutations inMAFB, a negative regulator of RANKL-induced osteoclastogenesis","volume":"164","author":"Mumm","year":"2014","journal-title":"Am. J. Med. Genet. Part A"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"494","DOI":"10.1016\/j.ajhg.2012.01.003","article-title":"Multicentric Carpotarsal Osteolysis Is Caused by Mutations Clustering in the Amino-Terminal Transcriptional Activation Domain of MAFB","volume":"90","author":"Zankl","year":"2012","journal-title":"Am. J. Hum. Genet."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Park, P.G., Kim, K.H., Hyun, H.S., Lee, C.H., Park, J.-S., Kie, J.H., Choi, Y.H., Moon, K.C., and Cheong, H.I. (2018). Three cases of multicentric carpotarsal osteolysis syndrome: A case series. BMC Med. Genet., 19.","DOI":"10.1186\/s12881-018-0682-x"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1016\/j.kint.2018.02.025","article-title":"A mutation in transcription factor MAFB causes Focal Segmental Glomerulosclerosis with Duane Retraction Syndrome","volume":"94","author":"Sato","year":"2018","journal-title":"Kidney Int."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"6783957","DOI":"10.1155\/2018\/6783957","article-title":"The First Report of Multicentric Carpotarsal Osteolysis Syndrome Caused by MAFB Mutation in Asian","volume":"2018","author":"Choochuen","year":"2018","journal-title":"Case Rep. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1538\/expanim.18-0093","article-title":"Mice harboring an MCTO mutation exhibit renal failure resembling nephropathy in human patients","volume":"68","author":"Tsunakawa","year":"2019","journal-title":"Exp. Anim."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/j.ydbio.2019.07.009","article-title":"Zebrafish and medaka as models for biomedical research of bone diseases","volume":"457","author":"Winkler","year":"2020","journal-title":"Dev. Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"6","DOI":"10.3389\/fendo.2019.00006","article-title":"Zebrafish as an Emerging Model for Osteoporosis: A Primary Testing Platform for Screening New Osteo-Active Compounds","volume":"10","author":"Bergen","year":"2019","journal-title":"Front. Endocrinol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1002\/jmor.1065","article-title":"Features of mono- and multinucleated bone resorbing cells of the zebrafishDanio rerio and their contribution to skeletal development, remodeling, and growth","volume":"250","author":"Witten","year":"2001","journal-title":"J. Morphol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"925","DOI":"10.1002\/jor.24539","article-title":"Zebrafish: An Emerging Model for Orthopedic Research","volume":"38","author":"Busse","year":"2019","journal-title":"J. Orthop. Res."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Gomez, G.A., Veldman, M.B., Zhao, Y., Burgess, S., and Lin, S. (2009). Discovery and Characterization of Novel Vascular and Hematopoietic Genes Downstream of Etsrp in Zebrafish. PLoS ONE, 4.","DOI":"10.1371\/journal.pone.0004994"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1016\/S0925-4773(98)00220-2","article-title":"Expression of Zkrml2, a homologue of the Krml1\/val segmentation gene, during embryonic patterning of the zebrafish (Danio rerio)","volume":"80","author":"Schvarzstein","year":"1999","journal-title":"Mech. Dev."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1093\/oxfordjournals.jbchem.a002825","article-title":"Isolation, characterization, and expression analysis of zebrafish large Mafs","volume":"129","author":"Kajihara","year":"2001","journal-title":"J. Biochem."},{"key":"ref_27","first-page":"1101","article-title":"Optimized CRISPR\u2013Cas9 System for Genome Editing in Zebrafish","volume":"10","author":"Vejnar","year":"2016","journal-title":"Cold Spring Harb. Protoc."},{"key":"ref_28","first-page":"211","article-title":"Osteoclast-like Cells in Early Zebrafish Embryos","volume":"16","author":"Sharif","year":"2014","journal-title":"Cell J."},{"key":"ref_29","first-page":"259","article-title":"In situ hybridization: A practical approach","volume":"35","author":"Wilkinson","year":"1992","journal-title":"Methods Cell Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1006\/dbio.2001.0393","article-title":"Zebrafish Early Macrophages Colonize Cephalic Mesenchyme and Developing Brain, Retina, and Epidermis through a M-CSF Receptor-Dependent Invasive Process","volume":"238","author":"Herbomel","year":"2001","journal-title":"Dev. Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1182\/blood.V26.2.215.215","article-title":"Simplified myeloperoxidase stain using benzidine dihydrochloride","volume":"26","author":"Kaplow","year":"1965","journal-title":"Blood"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/j.cbpa.2018.06.011","article-title":"RANKL, Ephrin-Eph and Wnt10b are key intercellular communication molecules regulating bone remodeling in au-tologous transplanted goldfish scales","volume":"225","author":"Tazaki","year":"2018","journal-title":"Comp. Biochem. Physiol. A Mol. Integr. Physiol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"100215","DOI":"10.1016\/j.bonr.2019.100215","article-title":"Aging-associated microstructural deterioration of vertebra in zebrafish","volume":"11","author":"Monma","year":"2019","journal-title":"Bone Rep."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1016\/j.bone.2017.05.001","article-title":"Utility of quantitative micro-computed tomographic analysis in zebrafish to define gene function during skeletogenesis","volume":"101","author":"Charles","year":"2017","journal-title":"Bone"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1080\/10520290701333558","article-title":"A two-color acid-free cartilage and bone stain for zebrafish larvae","volume":"82","author":"Walker","year":"2007","journal-title":"Biotech. Histochem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T))","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Chen, J.-R., Lai, Y.-H., Tsai, J.-J., and Hsiao, C.-D. (2017). Live Fluorescent Staining Platform for Drug-Screening and Mechanism-Analysis in Zebrafish for Bone Mineralization. Molecules, 22.","DOI":"10.20944\/preprints201709.0102.v1"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1016\/j.celrep.2017.06.070","article-title":"Robust Identification of Developmentally Active Endothelial Enhancers in Zebrafish Using FANS-Assisted ATAC-Seq","volume":"20","author":"Quillien","year":"2017","journal-title":"Cell Rep."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"e49","DOI":"10.1182\/blood-2010-10-314120","article-title":"mpeg1 promoter transgenes direct macrophage-lineage expression in zebrafish","volume":"117","author":"Ellett","year":"2011","journal-title":"Blood"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1242\/dev.071035","article-title":"Rankl-induced osteoclastogenesis leads to loss of mineralization in a medaka osteoporosis model","volume":"139","author":"To","year":"2011","journal-title":"Development"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"4016","DOI":"10.1096\/fj.201701424R","article-title":"OC-STAMP promotes osteoclast fusion for pathogenic bone resorption in periodontitis via up-regulation of permissive fusogen CD9","volume":"32","author":"Ishii","year":"2018","journal-title":"FASEB J."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1242\/dev.123.1.357","article-title":"Mutations affecting craniofacial development in zebrafish","volume":"123","author":"Neuhauss","year":"1996","journal-title":"Development"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.toxlet.2021.02.005","article-title":"A quantitative in vivo assay for craniofacial developmental toxicity of histone deacetylases","volume":"342","author":"Koch","year":"2021","journal-title":"Toxicol. Lett."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"R100","DOI":"10.1186\/ar3381","article-title":"Oestrogen deficiency modulates particle-induced osteolysis","volume":"13","author":"Nich","year":"2011","journal-title":"Arthritis Res."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1143","DOI":"10.1002\/jbmr.3689","article-title":"Cooperation of PU.1 With IRF8 and NFATc1 Defines Chromatin Landscapes During RANKL-Induced Osteoclasto-genesis","volume":"34","author":"Izawa","year":"2019","journal-title":"J. Bone Miner. Res."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"632","DOI":"10.1016\/j.devcel.2015.08.018","article-title":"Temporal-Spatial Resolution Fate Mapping Reveals Distinct Origins for Embryonic and Adult Microglia in Zebrafish","volume":"34","author":"Xu","year":"2015","journal-title":"Dev. Cell"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/S0092-8674(00)81081-8","article-title":"MafB Is an Interaction Partner and Repressor of Ets-1 That Inhibits Erythroid Differentiation","volume":"85","author":"Sieweke","year":"1996","journal-title":"Cell"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1055\/s-0038-1657760","article-title":"A Familial Case of Multicentric Carpotarsal Osteolysis Syndrome and Treatment Outcome","volume":"7","author":"Gomes","year":"2018","journal-title":"J. Pediatr. Genet."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1073\/pnas.96.1.133","article-title":"Alendronate mechanism of action: Geranylgeraniol, an intermediate in the mevalonate pathway, prevents inhibition of osteoclast formation, bone resorption, and kinase activation in vitro","volume":"96","author":"Fisher","year":"1999","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"2070","DOI":"10.4049\/jimmunol.1601667","article-title":"MAFB Determines Human Macrophage Anti-Inflammatory Polarization: Relevance for the Pathogenic Mechanisms Operating in Multicentric Carpotarsal Osteolysis","volume":"198","author":"Cuevas","year":"2017","journal-title":"J. Immunol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"867","DOI":"10.1126\/science.1176056","article-title":"MafB\/c-Maf Deficiency Enables Self-Renewal of Differentiated Functional Macrophages","volume":"326","author":"Aziz","year":"2009","journal-title":"Science"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1016\/j.cell.2009.04.057","article-title":"MafB Restricts M-CSF-Dependent Myeloid Commitment Divisions of Hematopoietic Stem Cells","volume":"138","author":"Sarrazin","year":"2009","journal-title":"Cell"}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/11\/3\/480\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:39:45Z","timestamp":1760161185000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/11\/3\/480"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,23]]},"references-count":52,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2021,3]]}},"alternative-id":["biom11030480"],"URL":"https:\/\/doi.org\/10.3390\/biom11030480","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,3,23]]}}}