{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,8]],"date-time":"2026-11-08T07:35:12Z","timestamp":1794123312603,"version":"4.8.0"},"reference-count":30,"publisher":"Oxford University Press (OUP)","issue":"5","license":[{"start":{"date-parts":[[2022,2,10]],"date-time":"2022-02-10T00:00:00Z","timestamp":1644451200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2022,5,18]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Malignant pleural effusion (MPE) represents advanced malignant disease with poor prognosis. To date, pleural effusion cytology remains the best test to diagnose MPE but suffers from limited diagnostic sensitivity and high variation. We report a hexokinase 2-based method (HK2-seq) as a novel diagnostic method for multicancer MPE diagnosis.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>HK2-seq employed HK2 as a new metabolic function-associated marker to detect disseminated tumor cells engaging increased glycolysis in pleural effusion from many cancer types. Single-cell sequencing was used to confirm the malignancy of HK2-derived high glycolytic tumor cells (hgTCs) at the single-cell level via surveying genome-wide copy number alterations (CNAs), leading to establishment of definitive MPE diagnosis.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>In a prospective cohort study including 111 patients with pleural effusion, the HK2 test showed diagnostic sensitivity, diagnostic specificity, positive predictive value, and negative predictive value of 91% (95% CI: 80%\u201397%), 84% (95% CI: 68%\u201393%), 90% (95% CI: 79%\u201396%), and 86% (95% CI: 70%\u201395%), respectively, in MPE diagnosis across 12 different cancer types. In contrast, pleural effusion cytology exhibits an overall diagnostic sensitivity of 45%. In addition to confirming the tumor origin of hgTCs, single-cell sequencing allowed identification of prognostic or targetable CNAs in hgTCs, especially CNAs found in liquid biopsies but absent in solid biopsies.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>HK2-seq establishes definitive MPE diagnosis across many cancer types with high diagnostic performance. It has the potential to be used for multicancer detection of circulating tumor cells in blood and other types of body fluids, as well as liquid biopsy-based genomic characterization for informative diagnosis.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1093\/clinchem\/hvac003","type":"journal-article","created":{"date-parts":[[2022,1,6]],"date-time":"2022-01-06T12:09:39Z","timestamp":1641470979000},"page":"680-690","source":"Crossref","is-referenced-by-count":13,"title":["A Multicancer Malignant Pleural Effusion Diagnostic Test Using Hexokinase 2 and Single-Cell Sequencing"],"prefix":"10.1093","volume":"68","author":[{"given":"Jie","family":"Chen","sequence":"first","affiliation":[{"name":"Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yibin","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Pathology, The Affiliated Hospital of Hangzhou Normal University, Hangzhou, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhuo","family":"Wang","sequence":"additional","affiliation":[{"name":"Shanghai Key Laboratory of Medical Epigenetics, International Co-Laboratory of Medical Epigenetics and Metabolism (Ministry of Science and Technology), Institutes of Biomedical Sciences, Fudan University, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohan","family":"Shen","sequence":"additional","affiliation":[{"name":"Shanghai Key Laboratory of Medical Epigenetics, International Co-Laboratory of Medical Epigenetics and Metabolism (Ministry of Science and Technology), Institutes of Biomedical Sciences, Fudan University, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziyuan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Whole-Period Monitoring and Precise Intervention of Digestive Cancer (SMHC), Institute of Fudan-Minhang Academic Health System, Minhang Hospital, Fudan University, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunying","family":"Wang","sequence":"additional","affiliation":[{"name":"Shanghai Key Laboratory of Medical Epigenetics, International Co-Laboratory of Medical Epigenetics and Metabolism (Ministry of Science and Technology), Institutes of Biomedical Sciences, Fudan University, Shanghai, China"},{"name":"Key Laboratory of Whole-Period Monitoring and Precise Intervention of Digestive Cancer (SMHC), Institute of Fudan-Minhang Academic Health System, Minhang Hospital, Fudan University, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haimiao","family":"Xu","sequence":"additional","affiliation":[{"name":"Department of Pathology, Zhejiang Cancer Hospital, Hangzhou, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8403-9452","authenticated-orcid":false,"given":"Qihui","family":"Shi","sequence":"additional","affiliation":[{"name":"Shanghai Key Laboratory of Medical Epigenetics, International Co-Laboratory of Medical Epigenetics and Metabolism (Ministry of Science and Technology), Institutes of Biomedical Sciences, Fudan University, Shanghai, China"},{"name":"Key Laboratory of Whole-Period Monitoring and Precise Intervention of Digestive Cancer (SMHC), Institute of Fudan-Minhang Academic Health System, Minhang Hospital, Fudan University, Shanghai, China"},{"name":"Shanghai Engineering Research Center of Biomedical Analysis Reagents, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2022,2,10]]},"reference":[{"key":"2022051823121613600_hvac003-B1","doi-asserted-by":"crossref","first-page":"1971","DOI":"10.1056\/NEJMcp010731","article-title":"Clinical practice. Pleural effusion","volume":"346","author":"Light","year":"2002","journal-title":"N Engl J Med"},{"key":"2022051823121613600_hvac003-B2","doi-asserted-by":"crossref","first-page":"1098","DOI":"10.1136\/thoraxjnl-2014-205285","article-title":"Predicting survival in malignant pleural effusion: development and validation of the LENT prognostic score","volume":"69","author":"Clive","year":"2014","journal-title":"Thorax"},{"issue":"suppl 2","key":"2022051823121613600_hvac003-B3","doi-asserted-by":"crossref","first-page":"ii4","DOI":"10.1136\/thx.2010.136978","article-title":"Investigation of a unilateral pleural effusion in adults: British Thoracic Society Pleural Disease Guideline 2010","volume":"65","author":"Hooper","year":"2010","journal-title":"Thorax"},{"key":"2022051823121613600_hvac003-B4","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.ccm.2012.11.002","article-title":"Pleural fluid biomarkers: beyond the Light criteria","volume":"34","author":"Porcel","year":"2013","journal-title":"Clin Chest Med"},{"key":"2022051823121613600_hvac003-B5","doi-asserted-by":"crossref","first-page":"582","DOI":"10.1513\/AnnalsATS.201303-068OC","article-title":"Use of circulating tumor cell technology (CELLSEARCH) for the diagnosis of malignant pleural effusions","volume":"10","author":"Lustgarten","year":"2013","journal-title":"Ann Am Thorac Soc"},{"key":"2022051823121613600_hvac003-B6","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.lungcan.2018.11.020","article-title":"Measurement and immunophenotyping of pleural fluid EpCAM-positive cells and clusters for the management of non-small cell lung cancer patients","volume":"127","author":"Thompson","year":"2019","journal-title":"Lung Cancer"},{"key":"2022051823121613600_hvac003-B7","doi-asserted-by":"crossref","first-page":"1405","DOI":"10.1097\/00000478-200111000-00008","article-title":"The role of desmin and N-cadherin in effusion cytology: a comparative study using established markers of mesothelial and epithelial cells","volume":"25","author":"Davidson","year":"2001","journal-title":"Am J Surg Pathol"},{"key":"2022051823121613600_hvac003-B8","doi-asserted-by":"crossref","first-page":"e2012228118","DOI":"10.1073\/pnas.2012228118","article-title":"Hexokinase 2 discerns a novel circulating tumor cell population associated with poor prognosis in lung cancer patients","volume":"118","author":"Yang","year":"2021","journal-title":"Proc Natl Acad Sci USA"},{"key":"2022051823121613600_hvac003-B9","doi-asserted-by":"crossref","first-page":"16284","DOI":"10.1021\/acs.analchem.0c04282","article-title":"Single-cell sequencing-enabled hexokinase 2 assay for noninvasive bladder cancer diagnosis and screening by detecting rare malignant cells in urine","volume":"92","author":"Wang","year":"2020","journal-title":"Anal Chem"},{"key":"2022051823121613600_hvac003-B10","doi-asserted-by":"crossref","first-page":"3856","DOI":"10.1038\/s41467-019-11808-3","article-title":"Liquid biopsy-based single-cell metabolic phenotyping of lung cancer patients for informative diagnostics","volume":"10","author":"Li","year":"2019","journal-title":"Nat Commun"},{"key":"2022051823121613600_hvac003-B11","doi-asserted-by":"crossref","first-page":"4777","DOI":"10.1038\/sj.onc.1209603","article-title":"Hexokinase II: cancer's double-edged sword acting as both facilitator and gatekeeper of malignancy when bound to mitochondria","volume":"25","author":"Mathupala","year":"2006","journal-title":"Oncogene"},{"key":"2022051823121613600_hvac003-B12","doi-asserted-by":"crossref","first-page":"43407","DOI":"10.1074\/jbc.M108181200","article-title":"Glucose catabolism in cancer cells: identification and characterization of a marked activation response of the type II hexokinase gene to hypoxic conditions","volume":"276","author":"Mathupala","year":"2001","journal-title":"J Biol Chem"},{"key":"2022051823121613600_hvac003-B13","author":"National Genomics Data Center"},{"key":"2022051823121613600_hvac003-B14","doi-asserted-by":"crossref","first-page":"1134","DOI":"10.1038\/ng.2760","article-title":"Pan-cancer patterns of somatic copy number alteration","volume":"45","author":"Zack","year":"2013","journal-title":"Nat Genet"},{"key":"2022051823121613600_hvac003-B15","doi-asserted-by":"crossref","first-page":"714","DOI":"10.1101\/gr.188060.114","article-title":"Optimizing sparse sequencing of single cells for highly multiplex copy number profiling","volume":"25","author":"Baslan","year":"2015","journal-title":"Genome Res"},{"key":"2022051823121613600_hvac003-B16","doi-asserted-by":"crossref","first-page":"11139","DOI":"10.1073\/pnas.1411446111","article-title":"Copy number alteration burden predicts prostate cancer relapse","volume":"111","author":"Hieronymus","year":"2014","journal-title":"Proc Natl Acad Sci USA"},{"key":"2022051823121613600_hvac003-B17","doi-asserted-by":"crossref","first-page":"e37294","DOI":"10.7554\/eLife.37294","article-title":"Tumor copy number alteration burden is a pan-cancer prognostic factor associated with recurrence and death","volume":"7","author":"Hieronymus","year":"2018","journal-title":"Elife"},{"key":"2022051823121613600_hvac003-B18","doi-asserted-by":"crossref","first-page":"421","DOI":"10.3389\/fgene.2018.00421","article-title":"Association analysis of somatic copy number alteration burden with breast cancer survival","volume":"9","author":"Zhang","year":"2018","journal-title":"Front Genet"},{"key":"2022051823121613600_hvac003-B19","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1038\/nm.4239","article-title":"Molecular analysis of circulating tumor cells identifies distinct copy-number profiles in patients with chemosensitive and chemorefractory small-cell lung cancer","volume":"23","author":"Carter","year":"2017","journal-title":"Nat Med"},{"key":"2022051823121613600_hvac003-B20","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/j.cell.2018.03.022","article-title":"Cell-of-origin patterns dominate the molecular classification of 10,000 tumors from 33 types of cancer","volume":"173","author":"Hoadley","year":"2018","journal-title":"Cell"},{"key":"2022051823121613600_hvac003-B21","doi-asserted-by":"crossref","first-page":"697","DOI":"10.1038\/nature14418","article-title":"TP53 loss creates therapeutic vulnerability in colorectal cancer","volume":"520","author":"Liu","year":"2015","journal-title":"Nature"},{"key":"2022051823121613600_hvac003-B22","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.cell.2012.03.003","article-title":"MYC on the path to cancer","volume":"149","author":"Dang","year":"2012","journal-title":"Cell"},{"key":"2022051823121613600_hvac003-B23","doi-asserted-by":"crossref","first-page":"938","DOI":"10.1038\/modpathol.2012.36","article-title":"Intratumoral heterogeneity of HER2 gene amplification in breast cancer: its clinicopathological significance","volume":"25","author":"Seol","year":"2012","journal-title":"Mod Pathol"},{"key":"2022051823121613600_hvac003-B24","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1038\/nature12628","article-title":"Selection and adaptation during metastatic cancer progression","volume":"501","author":"Klein","year":"2013","journal-title":"Nature"},{"key":"2022051823121613600_hvac003-B25","doi-asserted-by":"crossref","first-page":"719","DOI":"10.1038\/s41422-018-0044-4","article-title":"MAP3K1 and MAP2K4 mutations are associated with sensitivity to MEK inhibitors in multiple cancer models","volume":"28","author":"Xue","year":"2018","journal-title":"Cell Res"},{"key":"2022051823121613600_hvac003-B26","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1038\/s41523-019-0126-6","article-title":"PIK3CA and MAP3K1 alterations imply luminal A status and are associated with clinical benefit from pan-PI3K inhibitor buparlisib and letrozole in ER+ metastatic breast cancer","volume":"5","author":"Nixon","year":"2019","journal-title":"NPJ Breast Cancer"},{"key":"2022051823121613600_hvac003-B27","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1007\/s10549-019-05320-x","article-title":"Somatic loss of PIK3R1 may sensitize breast cancer to inhibitors of the MAPK pathway","volume":"177","author":"Turturro","year":"2019","journal-title":"Breast Cancer Res Treat"},{"key":"2022051823121613600_hvac003-B28","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1056\/NEJMoa1113205","article-title":"Intratumor heterogeneity and branched evolution revealed by multiregion sequencing","volume":"366","author":"Gerlinger","year":"2012","journal-title":"N Engl J Med"},{"key":"2022051823121613600_hvac003-B29","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1373\/clinchem.2019.303586","article-title":"Molecular and functional characterization of circulating tumor cells: from discovery to clinical application","volume":"66","author":"Cort\u00e9s-Hern\u00e1ndez","year":"2020","journal-title":"Clin Chem"},{"key":"2022051823121613600_hvac003-B30","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1038\/s41568-019-0180-2","article-title":"Unravelling tumour heterogeneity by single-cell profiling of circulating tumour cells","volume":"19","author":"Keller","year":"2019","journal-title":"Nat Rev Cancer"}],"container-title":["Clinical Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/clinchem\/advance-article-pdf\/doi\/10.1093\/clinchem\/hvac003\/42489570\/hvac003.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/clinchem\/article-pdf\/68\/5\/680\/43752698\/hvac003.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/clinchem\/article-pdf\/68\/5\/680\/43752698\/hvac003.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,5,18]],"date-time":"2022-05-18T23:13:38Z","timestamp":1652915618000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/clinchem\/article\/68\/5\/680\/6526183"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,10]]},"references-count":30,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2022,2,10]]},"published-print":{"date-parts":[[2022,5,18]]}},"URL":"https:\/\/doi.org\/10.1093\/clinchem\/hvac003","relation":{},"ISSN":["0009-9147","1530-8561"],"issn-type":[{"value":"0009-9147","type":"print"},{"value":"1530-8561","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2022,5,3]]},"published":{"date-parts":[[2022,2,10]]}}}