Personalized analysis of minimal residual cancer cells in peritoneal lavage fluid predicts peritoneal dissemination of gastric cancer

Personalized analysis of minimal residual cancer cells in peritoneal lavage fluid predicts peritoneal dissemination of gastric cancer | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Personalized analysis of minimal residual cancer cells in peritoneal lavage fluid predicts peritoneal dissemination of gastric cancer Dongbin Zhao, Pinli Yue, Tongbo Wang, Pei Wang, Qianqian Song, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-275520/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Peritoneal dissemination (PD) is the major type of gastric cancer (GC) recurrence and leads to rapid death. Current approach cannot precisely determine which patients are at high risk of PD. In this study, we developed a technology to detect minimal residual cancer cells in peritoneal lavage fluid (PLF) samples by parallel profiling tumor-specific mutations. We applied the technology to a prospective cohort of 110 GC patients. The technology showed ultra-high sensitivity by successfully detecting all the 27 cases that developed PD. The minimal residual cancer cells in PLF was associated with an increased risk of PD (HR = 145.13; 95%CI = 20.20-18435.79; p < 0.001) and significantly shorter overall survival. In pathologically high-risk (T4) patients, the PLF mutation profiling model exhibited even greater specificity of 91% and positive predictive value of 88%, while retaining sensitivity of 100%. PLF cancer cell fraction remained the strongest independent predictor of PD and recurrence-free survival over pathologic diagnosis and cytological diagnosis in GC patients. This approach may help in the postsurgical management of GC patients by detecting PD far before the metastatic lesions grow to significant size detectable by conventional methods such as MRI and CT scanning. Cancer Biology Oncology Gastric Cancer Peritoneal lavage fluid Peritoneal dissemination Personalized mutation assay Minimal residual disease Figures Figure 1 Figure 2 Figure 3 Figure 4 Full Text Due to technical limitations, full-text HTML conversion of this manuscript could not be completed. However, the latest manuscript can be downloaded and accessed as a PDF. Additional Declarations Yes there is potential Competing Interest. Y.J. is one of the cofounders and have owner interest in Genetron Holdings, and receives royalties from Genetron. Y.J., D.Z., P.W., Q.S., and P.Y. have filed patents/patent applications based on the technology and data generated from this work. The remaining authors disclose no conflicts. Supplementary Files SupplementaryFigures.pdf Supplementary Figures SupplementaryTables.xlsx Supplementary Tables Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-275520","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":13659623,"identity":"3de99851-443f-40ec-aaa8-e6fff6f191e8","order_by":0,"name":"Dongbin Zhao","email":"","orcid":"","institution":"Chinese Academy of Medical Sciences and Peking Union Medical College","correspondingAuthor":false,"prefix":"","firstName":"Dongbin","middleName":"","lastName":"Zhao","suffix":""},{"id":13659624,"identity":"967053a5-edc2-47c4-b393-8fc487af8ce6","order_by":1,"name":"Pinli Yue","email":"","orcid":"","institution":"Chinese Academy of Medical Sciences and Peking Union Medical College","correspondingAuthor":false,"prefix":"","firstName":"Pinli","middleName":"","lastName":"Yue","suffix":""},{"id":13659625,"identity":"3f8b3d95-d6f9-42e1-8a89-fcb8d1b2f15a","order_by":2,"name":"Tongbo Wang","email":"","orcid":"","institution":"National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College","correspondingAuthor":false,"prefix":"","firstName":"Tongbo","middleName":"","lastName":"Wang","suffix":""},{"id":13659626,"identity":"0f104b04-96fa-413e-bcd7-670858a7abe7","order_by":3,"name":"Pei Wang","email":"","orcid":"","institution":"Chinese Academy of Medical Sciences and Peking Union Medical College","correspondingAuthor":false,"prefix":"","firstName":"Pei","middleName":"","lastName":"Wang","suffix":""},{"id":13659627,"identity":"9824ba15-4c49-4725-91ff-9f327abc86bf","order_by":4,"name":"Qianqian Song","email":"","orcid":"","institution":"Chinese Academy of Medical Sciences and Peking Union Medical College","correspondingAuthor":false,"prefix":"","firstName":"Qianqian","middleName":"","lastName":"Song","suffix":""},{"id":13659628,"identity":"bd973822-7ae6-4b87-9825-93cd03d8cc5f","order_by":5,"name":"Jingjing Wang","email":"","orcid":"","institution":"Jinchenjunchuang Clinical Laboratory","correspondingAuthor":false,"prefix":"","firstName":"Jingjing","middleName":"","lastName":"Wang","suffix":""},{"id":13659629,"identity":"da714f9a-e8a6-4562-983c-7aab882f71bc","order_by":6,"name":"Yuchen jiao","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzklEQVRIiWNgGAWjYBACxmYIncDAwHzgwIcK0rSwJR6ccYYE24BaeIwP87YQoZS5nfnhY942uzx+6Z4PB3gbGOT5xQ4QchibsTFvW3Kx5JyzGw5I7mAwnDk7gZAWBjNp3rYDiRtu5G44YHiGIcHgNkEt7N/AWvbfyHlwILGNKC08UFskchgOHCRSS7HhnHPJiTNupBkcbDgjQdgvhv3HNz54U2aX2D8j+fHnPxU28vzShLQ0MDAw8SD4EviVg4A8yHE/CKsbBaNgFIyCkQwApwlIpKidcrAAAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0001-7917-8810","institution":"Chinese Academy of Medical Sciences and Peking Union Medical College","correspondingAuthor":true,"prefix":"","firstName":"Yuchen","middleName":"","lastName":"jiao","suffix":""}],"badges":[],"createdAt":"2021-02-25 06:25:34","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-275520/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-275520/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":6805887,"identity":"2662149a-8463-477c-80ae-9183e48ee157","added_by":"auto","created_at":"2021-03-10 17:20:12","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":68004,"visible":true,"origin":"","legend":"Patient enrollment, sample detection workflow and the prognosis prediction of patients.","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-275520/v1/60df55654500db108504b04e.png"},{"id":6805890,"identity":"1471795b-61ee-4697-9839-10e152e92c04","added_by":"auto","created_at":"2021-03-10 17:20:13","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":80321,"visible":true,"origin":"","legend":"Cancer cell fraction model and background noise. (a). Cancer cell fraction model. A model to estimate the cancer cell fraction based on allele frequency and sequencing depth of somatic mutations in tumor tissue and paired PLF samples. MAF: mutant allele frequency; 𝑃t𝑖: MAF in solid tumor tissue; 𝑃c𝑖: MAF in corresponding peritoneal lavage fluid (PLF); 𝐷𝑖: sequencing depth in PLF; 𝐴𝑖: mutation reads number in PLF; 𝑋𝑖: observing reads with mutation in PLF; and R: overall cancer cell concentration. \n(b). The linear correlation between theoretical and estimated cancer cell fraction up to the dilution of 1:10-5. Each dilution was repeated three times. The blue dots highlight the fractions above the limit of detection (PLC/PRF/5 cell fraction = 0.001%, 0.005%, 0.05%, 0.5%, 5% and 33%). The red dots highlight the fractions under the limit of detection (PLC/PRF/5 cell fraction = 0.0001%, 0.0003%). \n(c). Background noise observed in the cancer cell fraction model at 0% PLC/PRF/5 cell input among the 20 independent replicates (green dots). Experiments performed on different cell line dilutions (0.0001% and 0.0003%, red dots; 0.001%, blue dots) were repeated three times. \n(d). Biological noise of the 104 PLF samples from patients. The cancer cell fraction for each sample was calculated based on non-tumor-specific mutations.","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-275520/v1/243628bf66f134facebc5c48.png"},{"id":6806219,"identity":"ebd4c5b3-4bfe-406c-9be9-a2b03cf07a2d","added_by":"auto","created_at":"2021-03-10 17:23:13","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":74737,"visible":true,"origin":"","legend":"Summary of clinical and histopathologic parameters, somatic mutations and cancer cell fraction for all patients. (a). Top panel, the summary of the frequencies of the tracked mutations in tumor and matched peritoneal lavage fluid samples from 104 patients. Blue bar, the tumor frequency of each tracked mutation. Frequency values are shown on the left vertical axis. Red bar, the detected peritoneal lavage fluid frequency of each tracked mutation. Frequency values are shown on the right vertical axis. The clinical outcome of patients is indicated under the bar. Middle panel, the summary of the cancer cell fractions for each patient. Bottom panel, clinical and histopathological characteristics. \n(b). The cancer cell fraction distribution in patients with peritoneal dissemination (n = 27), lymphatic metastasis (n = 6) or no recurrence (n = 71). Reported p values are computed using 2-tailed Wilcoxon Mann-Whitney U test.\n***, p \u003c 0.001; *, p \u003c 0.05","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-275520/v1/76edd850b85c475975fa945f.png"},{"id":6806220,"identity":"f050ae36-9bf5-46c0-872e-3316af7fe734","added_by":"auto","created_at":"2021-03-10 17:23:13","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":45468,"visible":true,"origin":"","legend":"Kaplan-Meier estimates of recurrence-free survival (RFS) and overall survival (OS) for gastric cancer patients. (a) and (b). Kaplan-Meier survival analysis shows probability of recurrence-free survival (RFS) as determined by A. cancer cell fraction detected in peritoneal lavage fluid (n = 98); and B. cancer cell fraction detected in peritoneal lavage fluid in stage T4 patients (n = 56) (for peritoneal dissemination). A patient was classified as testing positive if the cancer cell fraction detected in peritoneal lavage fluid was \u003e 0.01%; \n(c) Kaplan-Meier estimates of overall survival (OS) for 104 gastric cancer patients based on the estimated cancer cell fraction in peritoneal lavage fluid. \nShaded areas in the Kaplan-Meier plots indicate 95%CIs. HR: hazard ratio; CCF: cancer cell fraction.","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-275520/v1/fe2fbff11fed7bc3eaf356ae.png"},{"id":13602700,"identity":"13655bba-3ad0-47c1-9c28-346de16b3b8b","added_by":"auto","created_at":"2021-09-17 05:52:30","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1233174,"visible":true,"origin":"","legend":"","description":"","filename":"maindocument.pdf","url":"https://assets-eu.researchsquare.com/files/rs-275520/v1_covered.pdf"},{"id":6806287,"identity":"ec75b060-15c6-4ed3-8f53-91228cb1105b","added_by":"auto","created_at":"2021-03-10 17:26:24","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1032210,"visible":true,"origin":"","legend":"","description":"","filename":"maindocument.pdf","url":"https://assets-eu.researchsquare.com/files/rs-275520/v1_stamped.pdf"},{"id":6806218,"identity":"36731bf1-434a-44a9-98dd-9dc08fbb4c68","added_by":"auto","created_at":"2021-03-10 17:23:12","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":421000,"visible":true,"origin":"","legend":"Supplementary Figures","description":"","filename":"SupplementaryFigures.pdf","url":"https://assets-eu.researchsquare.com/files/rs-275520/v1/7d15e659f07bb47642492f97.pdf"},{"id":6805892,"identity":"13c5f4d2-6fdd-4fef-a942-ef2aba0947f0","added_by":"auto","created_at":"2021-03-10 17:20:13","extension":"xlsx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":233192,"visible":true,"origin":"","legend":"Supplementary Tables","description":"","filename":"SupplementaryTables.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-275520/v1/17959549158944ff2f6d41bf.xlsx"}],"financialInterests":"\u003cb\u003eYes\u003c/b\u003e there is potential Competing Interest.\nY.J. is one of the cofounders and have owner interest in Genetron Holdings, and receives royalties from Genetron. Y.J., D.Z., P.W., Q.S., and P.Y. have filed patents/patent applications based on the technology and data generated from this work. The remaining authors disclose no conflicts.","formattedTitle":"Personalized analysis of minimal residual cancer cells in peritoneal lavage fluid predicts peritoneal dissemination of gastric cancer","fulltext":[{"header":"Full Text","content":"Due to technical limitations, full-text HTML conversion of this manuscript could not be completed. However, the latest manuscript can be downloaded and \u003ca href='/article/rs-275520/latest.pdf' target='_blank'\u003e accessed as a PDF.\u003c/a\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Gastric Cancer, Peritoneal lavage fluid, Peritoneal dissemination, Personalized mutation assay, Minimal residual disease","lastPublishedDoi":"10.21203/rs.3.rs-275520/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-275520/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Peritoneal dissemination (PD) is the major type of gastric cancer (GC) recurrence and leads to rapid death. Current approach cannot precisely determine which patients are at high risk of PD. In this study, we developed a technology to detect minimal residual cancer cells in peritoneal lavage fluid (PLF) samples by parallel profiling tumor-specific mutations. We applied the technology to a prospective cohort of 110 GC patients. The technology showed ultra-high sensitivity by successfully detecting all the 27 cases that developed PD. The minimal residual cancer cells in PLF was associated with an increased risk of PD (HR = 145.13; 95%CI = 20.20-18435.79; p \u003c 0.001) and significantly shorter overall survival. In pathologically high-risk (T4) patients, the PLF mutation profiling model exhibited even greater specificity of 91% and positive predictive value of 88%, while retaining sensitivity of 100%. PLF cancer cell fraction remained the strongest independent predictor of PD and recurrence-free survival over pathologic diagnosis and cytological diagnosis in GC patients. This approach may help in the postsurgical management of GC patients by detecting PD far before the metastatic lesions grow to significant size detectable by conventional methods such as MRI and CT scanning.","manuscriptTitle":"Personalized analysis of minimal residual cancer cells in peritoneal lavage fluid predicts peritoneal dissemination of gastric cancer","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-03-10 17:20:11","doi":"10.21203/rs.3.rs-275520/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"6fdb066c-198d-4ee5-b741-9db47e3a60fc","owner":[],"postedDate":"March 10th, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":2873728,"name":"Cancer Biology"},{"id":2873729,"name":"Oncology"}],"tags":[],"updatedAt":"2021-04-12T11:25:41+00:00","versionOfRecord":[],"versionCreatedAt":"2021-03-10 17:20:11","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-275520","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-275520","identity":"rs-275520","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}