{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T05:19:20Z","timestamp":1790831960077,"version":"4.1.0"},"reference-count":40,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2022,3,4]],"date-time":"2022-03-04T00:00:00Z","timestamp":1646352000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["19ck0106259h0003"],"award-info":[{"award-number":["19ck0106259h0003"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["20ck0106541h0001"],"award-info":[{"award-number":["20ck0106541h0001"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["18H04030"],"award-info":[{"award-number":["18H04030"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["19H03498"],"award-info":[{"award-number":["19H03498"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007449","name":"Takeda Science Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100007449","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["pathsocjournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Journal of Pathology"],"published-print":{"date-parts":[[2022,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Loss\u2010of\u2010function mutations in <jats:italic>RNF43<\/jats:italic> induce activation of Wnt ligand\u2010dependent Wnt\/\u03b2\u2010catenin signaling through stabilization of the Frizzled receptor, which is often found in microsatellite instability (MSI)\u2010type colorectal cancer (CRC) that develops from sessile serrated adenomas. However, the mechanism underlying how <jats:italic>RNF43<\/jats:italic> mutations promote tumorigenesis remains poorly understood. In this study, we established nine human CRC\u2010derived organoids and found that three organoid lines carried <jats:italic>RNF43<\/jats:italic> frameshift mutations associated with MSI\u2010high and <jats:italic>BRAF<\/jats:italic><jats:sup><jats:italic>V600E<\/jats:italic><\/jats:sup> mutations, suggesting that these CRCs developed through the serrated pathway. <jats:italic>RNF43<\/jats:italic> frameshift mutant organoids required both Wnt ligands and R\u2010spondin for proliferation, indicating that suppression of ZNRF3 and retained RNF43 function by R\u2010spondin are required to achieve an indispensable level of Wnt activation for tumorigenesis. However, active \u03b2\u2010catenin levels in <jats:italic>RNF43<\/jats:italic>\u2010mutant organoids were lower than those in <jats:italic>APC<\/jats:italic> two\u2010hit mutant CRC, suggesting a lower threshold for Wnt activation in CRC that developed through the serrated pathway. Interestingly, transplantation of <jats:italic>RNF43<\/jats:italic>\u2010mutant organoids with intestinal myofibroblasts accelerated the \u03b2\u2010catenin nuclear accumulation and proliferation of xenograft tumors, indicating a key role of stromal cells in the promotion of the malignant phenotype of <jats:italic>RNF43<\/jats:italic>\u2010mutant CRC cells. Sequencing of subcloned organoid cell\u2010expressed transcripts revealed that two organoid lines carried monoallelic <jats:italic>RNF43 cis<\/jats:italic>\u2010mutations, with two <jats:italic>RNF43<\/jats:italic> frameshift mutations introduced in the same allele and the wild\u2010type <jats:italic>RNF43<\/jats:italic> allele remaining, while the other organoid line carried two\u2010hit biallelic <jats:italic>RNF43 trans<\/jats:italic>\u2010mutations. These results suggest that heterozygous <jats:italic>RNF43<\/jats:italic> frameshift mutations contribute to CRC development via the serrated pathway; however, a second\u2010hit <jats:italic>RNF43<\/jats:italic> mutation may be advantageous in tumorigenesis compared with a single\u2010hit mutation through further activation of Wnt signaling. Finally, treatment with the PORCN inhibitor significantly suppressed <jats:italic>RNF43<\/jats:italic>\u2010mutant cell\u2010derived PDX tumor development. These results suggest a novel mechanism underlying <jats:italic>RNF43<\/jats:italic> mutation\u2010associated CRC development and the therapeutic potential of Wnt ligand inhibition against <jats:italic>RNF43<\/jats:italic>\u2010mutant CRC. \u00a9 2022 The Authors. <jats:italic>The Journal of Pathology<\/jats:italic> published by John Wiley &amp; Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.<\/jats:p>","DOI":"10.1002\/path.5868","type":"journal-article","created":{"date-parts":[[2022,1,18]],"date-time":"2022-01-18T02:17:56Z","timestamp":1642472276000},"page":"39-52","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":40,"title":["Characterization of <scp><i>RNF43<\/i><\/scp> frameshift mutations that drive <scp>Wnt<\/scp> ligand\u2010 and <scp>R<\/scp>\u2010spondin\u2010dependent colon cancer"],"prefix":"10.1002","volume":"257","author":[{"given":"Daisuke","family":"Yamamoto","sequence":"first","affiliation":[{"name":"Division of Genetics, Cancer Research Institute Kanazawa University  Kanazawa Japan"},{"name":"Department of Thoracic, Cardiovascular and General Surgery Kanazawa University  Kanazawa Japan"},{"name":"Department of Gastroenterological Surgery Ishikawa Prefectural Central Hospital  Kanazawa Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6855-4008","authenticated-orcid":false,"given":"Hiroko","family":"Oshima","sequence":"additional","affiliation":[{"name":"Division of Genetics, Cancer Research Institute Kanazawa University  Kanazawa Japan"},{"name":"WPI Nano\u2010Life Science Institute (Nano\u2010LSI) Kanazawa University  Kanazawa Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2732-0808","authenticated-orcid":false,"given":"Dong","family":"Wang","sequence":"additional","affiliation":[{"name":"Division of Genetics, Cancer Research Institute Kanazawa University  Kanazawa Japan"},{"name":"WPI Nano\u2010Life Science Institute (Nano\u2010LSI) Kanazawa University  Kanazawa Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1643-2524","authenticated-orcid":false,"given":"Haruna","family":"Takeda","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Genetics, National Cancer Center Research Institute  Tokyo Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenji","family":"Kita","sequence":"additional","affiliation":[{"name":"Central Research Resource Branch, Cancer Research Institute Kanazawa University  Kanazawa Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuelian","family":"Lei","sequence":"additional","affiliation":[{"name":"Division of Genetics, Cancer Research Institute Kanazawa University  Kanazawa Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0464-3519","authenticated-orcid":false,"given":"Mizuho","family":"Nakayama","sequence":"additional","affiliation":[{"name":"Division of Genetics, Cancer Research Institute Kanazawa University  Kanazawa Japan"},{"name":"WPI Nano\u2010Life Science Institute (Nano\u2010LSI) Kanazawa University  Kanazawa Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1759-9542","authenticated-orcid":false,"given":"Kazuhiro","family":"Murakami","sequence":"additional","affiliation":[{"name":"Division of Stem Cell Biology, Cancer Research Institute Kanazawa University  Kanazawa Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takashi","family":"Ohama","sequence":"additional","affiliation":[{"name":"Laboratory of Veterinary Pharmacology, Joint Faculty of Veterinary Medicine Yamaguchi University  Yamaguchi Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hirofumi","family":"Takemura","sequence":"additional","affiliation":[{"name":"Department of Thoracic, Cardiovascular and General Surgery Kanazawa University  Kanazawa Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mutsumi","family":"Toyota","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology Sapporo Medical University School of Medicine  Sapporo Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9635-3238","authenticated-orcid":false,"given":"Hiromu","family":"Suzuki","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology Sapporo Medical University School of Medicine  Sapporo Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Noriyuki","family":"Inaki","sequence":"additional","affiliation":[{"name":"Department of Gastrointestinal Surgery Kanazawa University  Kanazawa Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3304-0004","authenticated-orcid":false,"given":"Masanobu","family":"Oshima","sequence":"additional","affiliation":[{"name":"Division of Genetics, Cancer Research Institute Kanazawa University  Kanazawa Japan"},{"name":"WPI Nano\u2010Life Science Institute (Nano\u2010LSI) Kanazawa University  Kanazawa Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2022,3,4]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2017.05.016"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature11308"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1508113112"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2017-315920"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng.3127"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1002\/path.4134"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng.2983"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1307218110"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/hgv.2015.13"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2016-311849"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2019.06.041"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.18632\/oncotarget.12130"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1097\/PAS.0000000000000664"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/path.4709"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2019-319126"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2018.03.035"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/path.5098"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng.3175"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1124\/mol.119.117978"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2015.03.053"},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/onc.2015.280"},{"key":"e_1_2_7_23_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-54931-3"},{"key":"e_1_2_7_24_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41388-020-1232-5"},{"key":"e_1_2_7_25_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-20-0957"},{"key":"e_1_2_7_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2017.12.009"},{"key":"e_1_2_7_27_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1223821"},{"key":"e_1_2_7_28_1","doi-asserted-by":"publisher","DOI":"10.3748\/wjg.v19.i17.2629"},{"key":"e_1_2_7_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajpath.2012.08.007"},{"key":"e_1_2_7_30_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13148-019-0668-3"},{"key":"e_1_2_7_31_1","doi-asserted-by":"publisher","DOI":"10.1093\/jnci\/djh034"},{"key":"e_1_2_7_32_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature11252"},{"key":"e_1_2_7_33_1","first-page":"5548","article-title":"Microsatellite instability and mutations of the transforming growth factor beta type II receptor gene in colorectal cancer","volume":"55","author":"Parsons R","year":"1995","journal-title":"Cancer Res"},{"key":"e_1_2_7_34_1","first-page":"994","article-title":"Evidence of selection for clones having genetic inactivation of the activin A type II receptor (ACVR2) gene in gastrointestinal cancers","volume":"63","author":"Hempen PM","year":"2003","journal-title":"Cancer Res"},{"key":"e_1_2_7_35_1","doi-asserted-by":"publisher","DOI":"10.1097\/PAS.0000000000000877"},{"key":"e_1_2_7_36_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature22334"},{"key":"e_1_2_7_37_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1713510115"},{"key":"e_1_2_7_38_1","doi-asserted-by":"publisher","DOI":"10.1002\/path.4087"},{"key":"e_1_2_7_39_1","first-page":"1644","article-title":"Morphological and molecular processes of polyp formation in Apc\u0394716 knockout mice","volume":"57","author":"Oshima H","year":"1997","journal-title":"Cancer Res"},{"key":"e_1_2_7_40_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1205822109"},{"key":"e_1_2_7_41_1","doi-asserted-by":"publisher","DOI":"10.1038\/nm.2772"}],"container-title":["The Journal of Pathology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/path.5868","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/path.5868","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/pathsocjournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/path.5868","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,21]],"date-time":"2025-10-21T16:34:17Z","timestamp":1761064457000},"score":1,"resource":{"primary":{"URL":"https:\/\/pathsocjournals.onlinelibrary.wiley.com\/doi\/10.1002\/path.5868"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,4]]},"references-count":40,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2022,5]]}},"alternative-id":["10.1002\/path.5868"],"URL":"https:\/\/doi.org\/10.1002\/path.5868","archive":["Portico"],"relation":{},"ISSN":["0022-3417","1096-9896"],"issn-type":[{"value":"0022-3417","type":"print"},{"value":"1096-9896","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,4]]},"assertion":[{"value":"2021-08-18","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2022-01-12","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2022-03-04","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}