{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,5]],"date-time":"2026-03-05T05:51:08Z","timestamp":1772689868015,"version":"3.50.1"},"reference-count":40,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2024,1,29]],"date-time":"2024-01-29T00:00:00Z","timestamp":1706486400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/100009619","name":"Japan Agency for Medical Research and Development","doi-asserted-by":"publisher","award":["JP23ama221410"],"award-info":[{"award-number":["JP23ama221410"]}],"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":["JP23fk0210110"],"award-info":[{"award-number":["JP23fk0210110"]}],"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":["JP23fk0210131"],"award-info":[{"award-number":["JP23fk0210131"]}],"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":["JP23fk0310501"],"award-info":[{"award-number":["JP23fk0310501"]}],"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":["JP23fk0310512"],"award-info":[{"award-number":["JP23fk0310512"]}],"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":["JP23fk0310524"],"award-info":[{"award-number":["JP23fk0310524"]}],"id":[{"id":"10.13039\/100009619","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cancer Science"],"published-print":{"date-parts":[[2024,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>There are approximately 250\u2009million people chronically infected with hepatitis B virus (HBV) worldwide. Although HBV is often integrated into the host genome and promotes hepatocarcinogenesis, vulnerability of HBV integration in liver cancer cells has not been clarified. The aim of our study is to identify vulnerability factors for HBV\u2010associated hepatocarcinoma. Loss\u2010of\u2010function screening was undertaken in HepG2 and HBV\u2010integrated HepG2.2.15 cells expressing SpCas9 using a pooled genome\u2010wide clustered regularly interspaced short palindromic repeats (CRISPR) library. Genes whose guide RNA (gRNA) abundance significantly decreased in HepG2.2.15 cells but not in HepG2 cells were extracted using the MAGeCK algorithm. We identified four genes (<jats:italic>BCL2L1<\/jats:italic>, <jats:italic>VPS37A<\/jats:italic>, <jats:italic>INSIG2<\/jats:italic>, and <jats:italic>CFLAR<\/jats:italic>) that showed significant reductions of gRNA abundance and thus potentially involved in the vulnerability of HBV\u2010integrated cancer cells. Among them, siRNA\u2010mediated mRNA inhibition or CRISPR\u2010mediated genetic deletion of <jats:italic>INSIG2<\/jats:italic> significantly impaired cell proliferation in HepG2.2.15 cells but not in HepG2 cells. Its inhibitory effect was alleviated by cotransfection of siRNAs targeting HBV. <jats:italic>INSIG2<\/jats:italic> inhibition suppressed the pathways related to cell cycle and DNA replication, downregulated cyclin\u2010dependent kinase 2 (<jats:italic>CDK2<\/jats:italic>) levels, and delayed the G<jats:sub>1<\/jats:sub>\u2010to\u2010S transition in HepG2.2.15 cells. <jats:italic>CDK2<\/jats:italic> inhibitor suppressed cell cycle progression in HepG2.2.15 cells and <jats:italic>INSIG2<\/jats:italic> inhibition did not suppress cell proliferation in the presence of <jats:italic>CDK2<\/jats:italic> inhibitor. In conclusion, <jats:italic>INSIG2<\/jats:italic> inhibition induced cell cycle arrest in HBV\u2010integrated hepatoma cells in a <jats:italic>CDK2<\/jats:italic>\u2010dependent manner, and thus <jats:italic>INSIG2<\/jats:italic> might be a vulnerability factor for HBV\u2010associated liver cancer.<\/jats:p>","DOI":"10.1111\/cas.16070","type":"journal-article","created":{"date-parts":[[2024,1,30]],"date-time":"2024-01-30T05:19:45Z","timestamp":1706591985000},"page":"859-870","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Genome\u2010wide loss\u2010of\u2010function genetic screen identifies <scp>INSIG2<\/scp> as the vulnerability of hepatitis B virus\u2010integrated hepatoma cells"],"prefix":"10.1111","volume":"115","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-6391-0201","authenticated-orcid":false,"given":"Makoto","family":"Fukuoka","sequence":"first","affiliation":[{"name":"Department of Gastroenterology and Hepatology Osaka University Graduate School of Medicine  Suita Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takahiro","family":"Kodama","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology and Hepatology Osaka University Graduate School of Medicine  Suita Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kazuhiro","family":"Murai","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology and Hepatology Osaka University Graduate School of Medicine  Suita Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hayato","family":"Hikita","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology and Hepatology Osaka University Graduate School of Medicine  Suita Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emi","family":"Sometani","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology and Hepatology Osaka University Graduate School of Medicine  Suita Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jihyun","family":"Sung","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology and Hepatology Osaka University Graduate School of Medicine  Suita Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akiyoshi","family":"Shimoda","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology and Hepatology Osaka University Graduate School of Medicine  Suita Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Satoshi","family":"Shigeno","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology and Hepatology Osaka University Graduate School of Medicine  Suita Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daisuke","family":"Motooka","sequence":"additional","affiliation":[{"name":"Genome Information Research Center, Research Institute for Microbial Diseases Osaka University  Suita Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akira","family":"Nishio","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology and Hepatology Osaka University Graduate School of Medicine  Suita Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kunimaro","family":"Furuta","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology and Hepatology Osaka University Graduate School of Medicine  Suita Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tomohide","family":"Tatsumi","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology and Hepatology Osaka University Graduate School of Medicine  Suita Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kosuke","family":"Yusa","sequence":"additional","affiliation":[{"name":"Stem Cell Genetics, Institute for Frontier Life and Medical Sciences Kyoto University  Kyoto Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tetsuo","family":"Takehara","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology and Hepatology Osaka University Graduate School of Medicine  Suita Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,1,29]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(14)60220-8"},{"key":"e_1_2_10_3_1","unstructured":"Organization WH.Guidelines for the prevention care and treatment of persons with chronic hepatitis B infection.2015."},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.2147\/JHC.S307962"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2020-323153"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41572-020-00240-3"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1225829"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/hep4.2014"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2800"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2016.09.079"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1705441114"},{"key":"e_1_2_10_12_1","first-page":"1183A","article-title":"Crispr loss\u2010of\u2010function screen identifies the hippo SIGNALLING pathway AS the mediator of regorafenib efficacy in hepatocellular carcinoma","volume":"70","author":"Kodama T","year":"2019","journal-title":"Hepatology"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.84.4.1005"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13073-021-00981-0"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-014-0554-4"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/hep.1840380410"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtn.2018.02.005"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2020.04.031"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00339-18"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molmet.2021.101428"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.46.29207"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1080\/15548627.2021.1917948"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20061091"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1158\/2159-8290.CD-20-1503"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/cdg210"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/s10038-017-0376-9"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-12606-7"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-03741-7"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-21-1017"},{"key":"e_1_2_10_30_1","first-page":"9","article-title":"Genome\u2010wide CRISPR\/Cas9 library screening revealed dietary restriction of glutamine in combination with inhibition of pyruvate metabolism as effective liver cancer treatment. Advanced","volume":"9","author":"Yang CX","year":"2022","journal-title":"Science"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2020.108676"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-18-3374"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.162488899"},{"key":"e_1_2_10_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2022.08.005"},{"key":"e_1_2_10_35_1","doi-asserted-by":"publisher","DOI":"10.2174\/1389202919666181002144336"},{"key":"e_1_2_10_36_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.8266103"},{"key":"e_1_2_10_37_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrc2602"},{"key":"e_1_2_10_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2005.12.022"},{"key":"e_1_2_10_39_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.24.11215"},{"key":"e_1_2_10_40_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.gene.2018.12.054"},{"key":"e_1_2_10_41_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng0195-41"}],"container-title":["Cancer Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/cas.16070","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,3,8]],"date-time":"2024-03-08T06:39:20Z","timestamp":1709879960000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/cas.16070"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,1,29]]},"references-count":40,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2024,3]]}},"alternative-id":["10.1111\/cas.16070"],"URL":"https:\/\/doi.org\/10.1111\/cas.16070","archive":["Portico"],"relation":{},"ISSN":["1347-9032","1349-7006"],"issn-type":[{"value":"1347-9032","type":"print"},{"value":"1349-7006","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,1,29]]},"assertion":[{"value":"2023-10-09","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-12-29","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-01-29","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}