{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T21:08:24Z","timestamp":1768511304239,"version":"3.49.0"},"posted":{"date-parts":[[2024,4,16]]},"group-title":"Cell Biology","reference-count":48,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,4,16]],"date-time":"2024-04-16T00:00:00Z","timestamp":1713225600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"name":"National Cancer Institute of the National Institutes of Health","award":["R37CA225807"],"award-info":[{"award-number":["R37CA225807"]}]},{"name":"National Institute of Diabetes and Digestive and Kidney Diseases of the National Institutes of Health","award":["P30DK078392"],"award-info":[{"award-number":["P30DK078392"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,5,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>As the role of hepatic progenitor cells (HPCs) constituting ductular reactions in pathogenesis remains ambiguous, we aimed to establish the in vivo cause-and-effect relationship between HPCs and chronic liver disease progression. We previously demonstrated that peritumoral ductules are associated with angiogenesis in liver tumors and forkhead box L1 (Foxl1)-expressing murine HPCs secrete angiogenic factors in vitro. Therefore, we hypothesized that HPCs are capable of remodeling the portal vascular microenvironment and regulating overall liver disease progression, and this function of HPCs is dependent on recombination signal binding protein for immunoglobulin kappa J region (RBPJ), a key effector of the Notch signaling pathway.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods and Results<\/jats:title>\n                  <jats:p>\n                    We generated HPC-specific\n                    <jats:italic>Rbpj<\/jats:italic>\n                    conditional knockout mice (CKO) using\n                    <jats:italic>Foxl1-Cre<\/jats:italic>\n                    and treated them with the DDC diet to induce chronic liver disease. CKO mice exhibited a significant reduction in serum levels of liver injury markers, ductular reactions, vascular and fibrotic areas, and hepatic expression of fibrosis and inflammation markers compared to control mice (WT). Single-nucleus RNA sequencing comparing CKO and WT livers detected transcriptome changes across multiple cell types, including endothelial cells, hepatic stellate cells, and cholangiocytes. Expression of several reactive cholangiocyte markers, including vascular cell adhesion molecule 1 (VCAM1), in HPCs was significantly downregulated in response to anti-\n                    <jats:italic>Rbpj<\/jats:italic>\n                    shRNAs in vitro. Immunofluorescence analysis indicated that the percentage of VCAM1+ cells was reduced in both HPC and cholangiocyte populations in CKO compared to WT in vivo.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>\n                    Our findings reveal\n                    <jats:italic>Rbpj<\/jats:italic>\n                    -dependent expression of reactive cholangiocyte markers in HPCs and demonstrate that\n                    <jats:italic>Rbpj<\/jats:italic>\n                    deletion in HPCs attenuates not only endothelial responses but also liver injury, fibrosis, and VCAM1 expression in cholangiocytes, highlighting the crucial role of HPCs in pathogenic progression.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2024.04.13.589277","type":"posted-content","created":{"date-parts":[[2024,4,16]],"date-time":"2024-04-16T08:40:18Z","timestamp":1713256818000},"source":"Crossref","is-referenced-by-count":0,"title":["<i>Rbpj<\/i>\n                  deletion in hepatic progenitor cells attenuates endothelial responses and fibrosis in DDC-fed mice"],"prefix":"10.64898","author":[{"given":"Sanghoon","family":"Lee","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lu","family":"Ren","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Weiwei","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aditi","family":"Paranjpe","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ping","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrew","family":"Potter","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stacey S.","family":"Huppert","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5696-9505","authenticated-orcid":false,"given":"Soona","family":"Shin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025050423500562000_2024.04.13.589277v2.1","doi-asserted-by":"publisher","DOI":"10.1002\/dvdy.22561"},{"key":"2025050423500562000_2024.04.13.589277v2.2","doi-asserted-by":"publisher","DOI":"10.1002\/hep.24613"},{"key":"2025050423500562000_2024.04.13.589277v2.3","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajpath.2019.04.011"},{"key":"2025050423500562000_2024.04.13.589277v2.4","doi-asserted-by":"publisher","DOI":"10.1002\/hep4.1204"},{"key":"2025050423500562000_2024.04.13.589277v2.5","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-416022-4.00010-X"},{"key":"2025050423500562000_2024.04.13.589277v2.6","doi-asserted-by":"crossref","unstructured":"Holczbauer A , Wangensteen KJ , Shin S . Cellular origins of regenerating liver and hepatocellular carcinoma. JHEP Rep 2022;4:100416.","DOI":"10.1016\/j.jhepr.2021.100416"},{"key":"2025050423500562000_2024.04.13.589277v2.7","first-page":"68","article-title":"Expression of vascular endothelial growth factors and their receptors by hepatic progenitor cells in human liver diseases","volume":"2","year":"2013","journal-title":"Hepatobiliary Surg Nutr"},{"key":"2025050423500562000_2024.04.13.589277v2.8","doi-asserted-by":"publisher","DOI":"10.1002\/hep.25742"},{"key":"2025050423500562000_2024.04.13.589277v2.9","doi-asserted-by":"publisher","DOI":"10.1101\/gad.2027811"},{"key":"2025050423500562000_2024.04.13.589277v2.10","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2014.09.039"},{"key":"2025050423500562000_2024.04.13.589277v2.11","doi-asserted-by":"publisher","DOI":"10.1002\/hep.28602"},{"key":"2025050423500562000_2024.04.13.589277v2.12","doi-asserted-by":"publisher","DOI":"10.2353\/ajpath.2007.061133"},{"key":"2025050423500562000_2024.04.13.589277v2.13","doi-asserted-by":"crossref","unstructured":"Lee S , Karns R , Shin S . Mechanism of paracrine communications between hepatic progenitor cells and endothelial cells. Cell Signal 2022;100:110458.","DOI":"10.1016\/j.cellsig.2022.110458"},{"key":"2025050423500562000_2024.04.13.589277v2.14","doi-asserted-by":"publisher","DOI":"10.1002\/hep.32299"},{"key":"2025050423500562000_2024.04.13.589277v2.15","doi-asserted-by":"publisher","DOI":"10.1016\/S1665-2681(19)31900-3"},{"key":"2025050423500562000_2024.04.13.589277v2.16","doi-asserted-by":"publisher","DOI":"10.1002\/hep.32140"},{"key":"2025050423500562000_2024.04.13.589277v2.17","doi-asserted-by":"publisher","DOI":"10.1042\/bj1800241"},{"key":"2025050423500562000_2024.04.13.589277v2.18","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0066094"},{"key":"2025050423500562000_2024.04.13.589277v2.19","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhep.2013.02.025"},{"key":"2025050423500562000_2024.04.13.589277v2.20","doi-asserted-by":"publisher","DOI":"10.1093\/intimm\/dxf030"},{"key":"2025050423500562000_2024.04.13.589277v2.21","doi-asserted-by":"publisher","DOI":"10.1002\/dvg.20315"},{"key":"2025050423500562000_2024.04.13.589277v2.22","doi-asserted-by":"crossref","unstructured":"Srinivas S , Watanabe T , Lin CS , William CM , Tanabe Y , Jessell TM , et al. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev Biol 2001;1:4.","DOI":"10.1186\/1471-213X-1-4"},{"key":"2025050423500562000_2024.04.13.589277v2.23","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2089"},{"key":"2025050423500562000_2024.04.13.589277v2.24","doi-asserted-by":"publisher","DOI":"10.1038\/nm.2667"},{"key":"2025050423500562000_2024.04.13.589277v2.25","doi-asserted-by":"publisher","DOI":"10.1016\/j.jtcme.2023.02.008"},{"key":"2025050423500562000_2024.04.13.589277v2.26","doi-asserted-by":"crossref","unstructured":"Byrnes K , Bailey NT , Baral K , Mercer A , Joshi S , Wahby N , et al. Impaired hepatic autophagy exacerbates hepatotoxin induced liver injury. Cell Death Discov 2023;9:71.","DOI":"10.1038\/s41420-023-01368-3"},{"key":"2025050423500562000_2024.04.13.589277v2.27","doi-asserted-by":"publisher","DOI":"10.1159\/000452214"},{"key":"2025050423500562000_2024.04.13.589277v2.28","doi-asserted-by":"crossref","unstructured":"Zhang Q , Wu J , Bai X , Liang T . Evaluation of Intra-Tumoral Vascularization in Hepatocellular Carcinomas. Front Med (Lausanne) 2020;7:584250.","DOI":"10.3389\/fmed.2020.584250"},{"key":"2025050423500562000_2024.04.13.589277v2.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhep.2019.04.012"},{"key":"2025050423500562000_2024.04.13.589277v2.30","doi-asserted-by":"publisher","DOI":"10.1002\/lt.21436"},{"key":"2025050423500562000_2024.04.13.589277v2.31","doi-asserted-by":"publisher","DOI":"10.1002\/hep.28948"},{"key":"2025050423500562000_2024.04.13.589277v2.32","doi-asserted-by":"publisher","DOI":"10.1016\/j.devcel.2022.01.004"},{"key":"2025050423500562000_2024.04.13.589277v2.33","doi-asserted-by":"crossref","unstructured":"MacParland SA , Liu JC , Ma XZ , Innes BT , Bartczak AM , Gage BK , et al. Single cell RNA sequencing of human liver reveals distinct intrahepatic macrophage populations. Nat Commun 2018;9:4383.","DOI":"10.1038\/s41467-018-06318-7"},{"key":"2025050423500562000_2024.04.13.589277v2.34","doi-asserted-by":"crossref","unstructured":"Li L , Pan H , Wang H , Li X , Bu X , Wang Q , et al. Interplay between VEGF and Nrf2 regulates angiogenesis due to intracranial venous hypertension. Sci Rep 2016;6:37338.","DOI":"10.1038\/srep37338"},{"key":"2025050423500562000_2024.04.13.589277v2.35","doi-asserted-by":"crossref","unstructured":"Chapouly C , Guimbal S , Hollier PL , Renault MA . Role of Hedgehog Signaling in Vasculature Development, Differentiation, and Maintenance. Int J Mol Sci 2019;20.","DOI":"10.3390\/ijms20123076"},{"key":"2025050423500562000_2024.04.13.589277v2.36","doi-asserted-by":"crossref","unstructured":"Cai X , Tacke F , Guillot A , Liu H . Cholangiokines: undervalued modulators in the hepatic microenvironment. Front Immunol 2023;14:1192840.","DOI":"10.3389\/fimmu.2023.1192840"},{"key":"2025050423500562000_2024.04.13.589277v2.37","doi-asserted-by":"publisher","DOI":"10.5009\/gnl.2012.6.3.295"},{"key":"2025050423500562000_2024.04.13.589277v2.38","doi-asserted-by":"publisher","DOI":"10.1002\/hep.26965"},{"key":"2025050423500562000_2024.04.13.589277v2.39","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0504391102"},{"key":"2025050423500562000_2024.04.13.589277v2.40","doi-asserted-by":"publisher","DOI":"10.1002\/stem.1030"},{"key":"2025050423500562000_2024.04.13.589277v2.41","doi-asserted-by":"crossref","unstructured":"Guo Q , Furuta K , Islam S , Caporarello N , Kostallari E , Dielis K , et al. Liver sinusoidal endothelial cell expressed vascular cell adhesion molecule 1 promotes liver fibrosis. Front Immunol 2022;13:983255.","DOI":"10.3389\/fimmu.2022.983255"},{"key":"2025050423500562000_2024.04.13.589277v2.42","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.20.2.648-655.2000"},{"key":"2025050423500562000_2024.04.13.589277v2.43","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtn.2021.11.008"},{"key":"2025050423500562000_2024.04.13.589277v2.44","doi-asserted-by":"crossref","unstructured":"Kong DH , Kim YK , Kim MR , Jang JH , Lee S . Emerging Roles of Vascular Cell Adhesion Molecule-1 (VCAM-1) in Immunological Disorders and Cancer. Int J Mol Sci 2018;19.","DOI":"10.3390\/ijms19041057"},{"key":"2025050423500562000_2024.04.13.589277v2.45","doi-asserted-by":"publisher","DOI":"10.1038\/cmi.2011.36"},{"key":"2025050423500562000_2024.04.13.589277v2.46","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvw207"},{"key":"2025050423500562000_2024.04.13.589277v2.47","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkac601"},{"key":"2025050423500562000_2024.04.13.589277v2.48","doi-asserted-by":"crossref","unstructured":"Martinez Lyons A , Boulter L . NOTCH signalling - a core regulator of bile duct disease? Dis Model Mech 2023;16.","DOI":"10.1242\/dmm.050231"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2024.04.13.589277","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T06:17:41Z","timestamp":1768457861000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2024.04.13.589277"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,4,16]]},"references-count":48,"URL":"https:\/\/doi.org\/10.1101\/2024.04.13.589277","relation":{"is-preprint-of":[{"id-type":"doi","id":"10.1097\/HC9.0000000000000745","asserted-by":"subject"}]},"subject":[],"published":{"date-parts":[[2024,4,16]]},"subtype":"preprint"}}