{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T01:42:46Z","timestamp":1760233366008,"version":"build-2065373602"},"reference-count":58,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2021,1,10]],"date-time":"2021-01-10T00:00:00Z","timestamp":1610236800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["NRF-2018R1A2B3005113","NRF-2018R1A5A2024418"],"award-info":[{"award-number":["NRF-2018R1A2B3005113","NRF-2018R1A5A2024418"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>CXC-chemokine receptor type 4 (CXCR4), a 7-transmembrane receptor family member, displays multifaceted roles, participating in immune cell migration, angiogenesis, and even adipocyte metabolism. However, the activity of such a ubiquitously expressed receptor in epithelial gland organogenesis has not yet been fully explored. To investigate the relationship between CXCL12\/CXCR4 signaling and embryonic glandular organogenesis, we used an ex vivo culture system with live imaging and RNA sequencing to elucidate the transcriptome and protein-level signatures of AMD3100, a potent abrogating reagent of the CXCR4-CXCL12 axis, imprinted on the developing organs. Immunostaining results showed that CXCR4 was highly expressed in embryonic submandibular gland, lung, and pancreas, especially at the periphery of end buds containing numerous embryonic stem\/progenitor cells. Despite no significant increase in apoptosis, AMD3100-treated epithelial organs showed a retarded growth with significantly slower branching and expansion. Further analyses with submandibular glands revealed that such responses resulted from the AMD3100-induced precocious differentiation of embryonic epithelial cells, losing mitotic activity. RNA sequencing analysis revealed that inhibition of CXCR4 significantly down-regulated polycomb repressive complex (PRC) components, known as regulators of DNA methylation. Treatment with PRC inhibitor recapitulated the AMD3100-induced precocious differentiation. Our results indicate that the epigenetic modulation by the PRC-CXCR12\/CXCR4 signaling axis is crucial for the spatiotemporal regulation of proliferation and differentiation of embryonic epithelial cells during embryonic glandular organogenesis.<\/jats:p>","DOI":"10.3390\/ijms22020619","type":"journal-article","created":{"date-parts":[[2021,1,10]],"date-time":"2021-01-10T19:55:56Z","timestamp":1610308556000},"page":"619","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["CXCR4 Regulates Temporal Differentiation via PRC1 Complex in Organogenesis of Epithelial Glands"],"prefix":"10.3390","volume":"22","author":[{"given":"Junchul","family":"Kim","sequence":"first","affiliation":[{"name":"Department of Physiology, School of Dentistry and Dental Research Institute, Seoul National University, Seoul 110-749, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sang-Woo","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Physiology, School of Dentistry and Dental Research Institute, Seoul National University, Seoul 110-749, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4908-1270","authenticated-orcid":false,"given":"Kyungpyo","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Physiology, School of Dentistry and Dental Research Institute, Seoul National University, Seoul 110-749, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,1,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1172\/JCI7954","article-title":"Chemotaxis of primitive hematopoietic cells in response to stromal cell\u2013derived factor-1","volume":"105","author":"Jo","year":"2000","journal-title":"J. Clin. Investig."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1925","DOI":"10.1073\/pnas.94.5.1925","article-title":"The HIV coreceptors CXCR4 and CCR5 are differentially expressed and regulated on human T lymphocytes","volume":"94","author":"Bleul","year":"1997","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"6165","DOI":"10.1038\/s41598-019-42127-8","article-title":"CXCL12-CXCR4 pathway activates brown adipocytes and induces insulin resistance in CXCR4-deficient mice under high-fat diet","volume":"9","author":"Kurita","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Pelekanos, R.A., Ting, M.J., Sardesai, V.S., Ryan, J.M., Lim, Y.-C., Chan, J.K.Y., and Fisk, N.M. (2014). Intracellular trafficking and endocytosis of CXCR4 in fetal mesenchymal stem\/stromal cells. BMC Cell Biol., 15.","DOI":"10.1186\/1471-2121-15-15"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1915","DOI":"10.1038\/sj.leu.2404357","article-title":"The pleiotropic effects of the SDF-1\u2013CXCR4 axis in organogenesis, regeneration and tumorigenesis","volume":"20","author":"Ratajczak","year":"2006","journal-title":"Leukemia"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"11","DOI":"10.7150\/thno.4806","article-title":"The role of CXCL12-CXCR4 signaling pathway in pancreatic development","volume":"3","author":"Katsumoto","year":"2013","journal-title":"Theranostics"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1080\/07357907.2017.1422512","article-title":"Knockdown of CXCR4 inhibits CXCL12-induced angiogenesis in HUVECs through downregulation of the MAPK\/ERK and PI3K\/AKT and the Wnt\/\u03b2-catenin pathways","volume":"36","author":"Song","year":"2018","journal-title":"Cancer Investig."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1714","DOI":"10.1681\/ASN.2008060640","article-title":"The CXCL12 (SDF-1)\/CXCR4 axis is essential for the development of renal vasculature","volume":"20","author":"Takabatake","year":"2009","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1038\/31261","article-title":"The chemokine receptor CXCR4 is essential for vascularization of the gastrointestinal tract","volume":"393","author":"Tachibana","year":"1998","journal-title":"Nature"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"LaRocca, T.J., Altman, P., Jarrah, A.A., Gordon, R., Wang, E., Hadri, L., Burke, M.W., Haddad, G.E., Hajjar, R.J., and Tarzami, S.T. (2019). CXCR4 cardiac specific knockout mice develop a progressive cardiomyopathy. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20092267"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1040","DOI":"10.1038\/labinvest.2011.77","article-title":"Targeted intestinal epithelial deletion of the chemokine receptor CXCR4 reveals important roles for extracellular-regulated kinase-1\/2 in restitution","volume":"91","author":"Zimmerman","year":"2011","journal-title":"Lab. Investig."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1387\/ijdb.140143dr","article-title":"Epithelial-mesenchymal interactions: A fundamental Developmental Biology mechanism","volume":"58","author":"Ribatti","year":"2014","journal-title":"Int. J. Dev. Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1186\/s13059-018-1416-2","article-title":"Single-cell RNA-seq analysis unveils a prevalent epithelial\/mesenchymal hybrid state during mouse organogenesis","volume":"19","author":"Dong","year":"2018","journal-title":"Genome Biol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Landsman, L., Nijagal, A., Whitchurch, T.J., Vanderlaan, R.L., Zimmer, W.E., MacKenzie, T.C., and Hebrok, M. (2011). Pancreatic mesenchyme regulates epithelial organogenesis throughout development. PLoS Biol., 9.","DOI":"10.1371\/journal.pbio.1001143"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4129","DOI":"10.1242\/dev.091793","article-title":"Sox proteins: Regulators of cell fate specification and differentiation","volume":"140","author":"Kamachi","year":"2013","journal-title":"Development"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.stem.2012.12.007","article-title":"The sox family of transcription factors: Versatile regulators of stem and progenitor cell fate","volume":"12","author":"Sarkar","year":"2013","journal-title":"Cell Stem Cell"},{"key":"ref_17","first-page":"194","article-title":"Signaling molecules governing pluripotency and early lineage commitments in human pluripotent stem cells","volume":"19","author":"Fathi","year":"2017","journal-title":"Cell J."},{"key":"ref_18","first-page":"2294","article-title":"Fgf10 and Sox9 are essential for the establishment of distal progenitor cells during mouse salivary gland development","volume":"144","author":"Chatzeli","year":"2017","journal-title":"Development"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1038\/s41596-018-0104-8","article-title":"Generation of lung organoids from human pluripotent stem cells in vitro","volume":"14","author":"Miller","year":"2019","journal-title":"Nat. Protoc."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1136\/gutjnl-2016-312423","article-title":"Human pluripotent stem cell-derived acinar\/ductal organoids generate human pancreas upon orthotopic transplantation and allow disease modelling","volume":"66","author":"Hohwieler","year":"2017","journal-title":"Gut"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/j.ejphar.2016.06.059","article-title":"Pluripotent stem cell-derived kidney organoids: An in vivo-like in vitro technology","volume":"790","author":"Schutgens","year":"2016","journal-title":"Eur. J. Pharmacol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4216","DOI":"10.1038\/s41467-018-06469-7","article-title":"Generation of orthotopically functional salivary gland from embryonic stem cells","volume":"9","author":"Tanaka","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2498","DOI":"10.1634\/stemcells.2006-0608","article-title":"Concise review: Roles of polycomb group proteins in development and disease: A stem cell perspective","volume":"25","author":"Rajasekhar","year":"2007","journal-title":"Stem Cells"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1042\/BST20160173","article-title":"Molecular architecture of polycomb repressive complexes","volume":"45","author":"Chittock","year":"2017","journal-title":"Biochem. Soc. Trans."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e1500737","DOI":"10.1126\/sciadv.1500737","article-title":"Regulation of gene transcription by Polycomb proteins","volume":"1","author":"Aranda","year":"2015","journal-title":"Sci. Adv."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1002\/wdev.4","article-title":"Salivary gland organogenesis","volume":"1","author":"Knosp","year":"2012","journal-title":"Wiley Interdiscip. Rev. Dev. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e26620","DOI":"10.7554\/eLife.26620","article-title":"SOX2 regulates acinar cell development in the salivary gland","volume":"6","author":"Emmerson","year":"2017","journal-title":"eLife"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1645","DOI":"10.1126\/science.1192046","article-title":"Parasympathetic innervation maintains epithelial progenitor cells during salivary organogenesis","volume":"329","author":"Knox","year":"2010","journal-title":"Science"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/j.actbio.2020.08.030","article-title":"Developmental role of hyaluronic acid and its application in salivary gland tissue engineering","volume":"115","author":"Lee","year":"2020","journal-title":"Acta Biomater."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1655","DOI":"10.1016\/j.bcp.2008.12.014","article-title":"The AMD3100 story: The path to the discovery of a stem cell mobilizer (Mozobil)","volume":"77","year":"2009","journal-title":"Biochem. Pharmacol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1060","DOI":"10.1038\/s41556-019-0384-4","article-title":"The cell cycle in stem cell proliferation, pluripotency and differentiation","volume":"21","author":"Liu","year":"2019","journal-title":"Nat. Cell Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1016\/j.devcel.2014.06.012","article-title":"Parasympathetic innervation regulates tubulogenesis in the developing salivary gland","volume":"30","author":"Nedvetsky","year":"2014","journal-title":"Dev. Cell"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"10827","DOI":"10.1038\/ncomms10827","article-title":"Mechanism of intermediate filament recognition by plakin repeat domains revealed by envoplakin targeting of vimentin","volume":"7","author":"Fogl","year":"2016","journal-title":"Nat. Commun."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2194","DOI":"10.1073\/pnas.0808602106","article-title":"Nesprin 4 is an outer nuclear membrane protein that can induce kinesin-mediated cell polarization","volume":"106","author":"Roux","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"808","DOI":"10.1016\/j.molcel.2013.02.013","article-title":"Occupying chromatin: Polycomb mechanisms for getting to genomic targets, stopping transcriptional traffic, and staying put","volume":"49","author":"Simon","year":"2013","journal-title":"Mol. Cell"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Endoh, M., Endo, T.A., Endoh, T., Isono, K.-i., Sharif, J., Ohara, O., Toyoda, T., Ito, T., Eskeland, R., and Bickmore, W.A. (2012). Histone H2A mono-ubiquitination is a crucial step to mediate PRC1-dependent repression of developmental genes to maintain ES cell identity. PLoS Genet., 8.","DOI":"10.1371\/journal.pgen.1002774"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"840","DOI":"10.1016\/j.molcel.2019.11.021","article-title":"Histone H2AK119 mono-ubiquitination is essential for polycomb-mediated transcriptional repression","volume":"77","author":"Tamburri","year":"2020","journal-title":"Mol. Cell"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Gertych, A., Oh, J.H., Wawrowsky, K.A., Weisenberger, D.J., and Tajbakhsh, J. (2013). 3-D DNA methylation phenotypes correlate with cytotoxicity levels in prostate and liver cancer cell models. BMC Pharmacol. Toxicol., 14.","DOI":"10.1186\/2050-6511-14-11"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1681","DOI":"10.4155\/fmc-2016-0082","article-title":"Structural aspects of small-molecule inhibition of methyllysine reader proteins","volume":"8","author":"Milosevich","year":"2016","journal-title":"Future Med. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"6085","DOI":"10.1182\/blood-2008-12-196618","article-title":"CXCR7 heterodimerizes with CXCR4 and regulates CXCL12-mediated G protein signaling","volume":"113","author":"Levoye","year":"2009","journal-title":"Blood"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1186\/1479-5876-10-68","article-title":"Chemokine CXCL12 activates dual CXCR4 and CXCR7-mediated signaling pathways in pancreatic cancer cells","volume":"10","author":"Heinrich","year":"2012","journal-title":"J. Transl. Med."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1897","DOI":"10.1038\/s41467-019-09810-w","article-title":"RUNX3 regulates cell cycle-dependent chromatin dynamics by functioning as a pioneer factor of the restriction-point","volume":"10","author":"Lee","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Hosogane, M., Funayama, R., Nishida, Y., Nagashima, T., and Nakayama, K. (2013). Ras-induced changes in H3K27me3 occur after those in transcriptional activity. PLoS Genet., 9.","DOI":"10.1371\/journal.pgen.1003698"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.gde.2015.01.007","article-title":"The contribution of specific cell subpopulations to submandibular salivary gland branching morphogenesis","volume":"32","author":"Kwon","year":"2015","journal-title":"Curr. Opin. Genet. Dev."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1800222","DOI":"10.1002\/bies.201800222","article-title":"Cell fate and developmental regulation dynamics by polycomb proteins and 3D genome architecture","volume":"41","author":"Loubiere","year":"2019","journal-title":"BioEssays"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"412","DOI":"10.1631\/jzus.B1400077","article-title":"Epigenetic regulation by polycomb group complexes: Focus on roles of CBX proteins","volume":"15","author":"Ma","year":"2014","journal-title":"J. Zhejiang Univ. Sci. B"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"2289","DOI":"10.1093\/nar\/gky1290","article-title":"Engagement of DNA and H3K27me3 by the CBX8 chromodomain drives chromatin association","volume":"47","author":"Connelly","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"3496","DOI":"10.1038\/s41467-019-11386-4","article-title":"Phc2 controls hematopoietic stem and progenitor cell mobilization from bone marrow by repressing Vcam1 expression","volume":"10","author":"Bae","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1513","DOI":"10.1242\/dev.014340","article-title":"Polycomb group proteins Ring1A\/B are functionally linked to the core transcriptional regulatory circuitry to maintain ES cell identity","volume":"135","author":"Endoh","year":"2008","journal-title":"Development"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"3135","DOI":"10.1038\/s41467-020-16919-w","article-title":"Genome-wide analysis in the mouse embryo reveals the importance of DNA methylation for transcription integrity","volume":"11","author":"Dahlet","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1089","DOI":"10.1126\/science.1063443","article-title":"Epigenetic reprogramming in mammalian development","volume":"293","author":"Reik","year":"2001","journal-title":"Science"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.devcel.2016.12.001","article-title":"Exosomal microRNA transport from salivary mesenchyme regulates epithelial progenitor expansion during organogenesis","volume":"40","author":"Hayashi","year":"2017","journal-title":"Dev. Cell"},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Shin, Y., Lee, S.-W., Namkoong, E., An, W., Lee, J.-H., Brown, P.D., and Park, K. (2019). Epigenetic modification as a regulatory mechanism for spatiotemporal dynamics of ANO1 expression in salivary glands. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20246298"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"7566","DOI":"10.1038\/s41598-018-25957-w","article-title":"Voltage-dependent Ca2+ channels promote branching morphogenesis of salivary glands by patterning differential growth","volume":"8","author":"Kim","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1016\/j.ydbio.2014.01.020","article-title":"DNA methylation of E-cadherin is a priming mechanism for prostate development","volume":"387","author":"Keil","year":"2014","journal-title":"Dev. Biol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"876","DOI":"10.1038\/nature01712","article-title":"Fibronectin requirement in branching morphogenesis","volume":"423","author":"Sakai","year":"2003","journal-title":"Nature"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"2927","DOI":"10.1158\/1078-0432.CCR-09-2329","article-title":"CXCL12 (SDF-1)\/CXCR4 pathway in cancer","volume":"16","author":"Teicher","year":"2010","journal-title":"Clin. Cancer Res."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"2109","DOI":"10.3389\/fimmu.2020.02109","article-title":"The chemokine receptor CXCR4 in cell proliferation and tissue regeneration","volume":"11","author":"Bianchi","year":"2020","journal-title":"Front. Immunol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/2\/619\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:09:17Z","timestamp":1760159357000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/2\/619"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,1,10]]},"references-count":58,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2021,1]]}},"alternative-id":["ijms22020619"],"URL":"https:\/\/doi.org\/10.3390\/ijms22020619","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2021,1,10]]}}}