Deconvolving metabolic intratumoral heterogeneity in clear cell renal carcinoma with hyperpolarized 13C-pyruvate MRI

Deconvolving metabolic intratumoral heterogeneity in clear cell renal carcinoma with hyperpolarized 13C-pyruvate MRI | 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 Research Article Deconvolving metabolic intratumoral heterogeneity in clear cell renal carcinoma with hyperpolarized 13 C-pyruvate MRI João André Gonçalves Duarte, Ines Horvat-Menih, Mary McLean, Stephan Ursprung, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7989219/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 9 You are reading this latest preprint version Abstract Background Clear cell renal cell carcinoma (ccRCC) is a highly heterogeneous cancer requiring a large number of biopsies to correctly characterize the tumor. Multiple biopsies are rarely feasible in the clinical setting, and therefore imaging methods offer the potential to evaluate the whole tumor non-invasively. For example, metabolic imaging offers the potential to probe the altered metabolism and metabolic heterogeneity that is characteristic of ccRCC. In this study we have explored and validated the use of hyperpolarized carbon-13 MRI (HP- 13 C-MRI) as a non-invasive clinical tool to probe metabolic heterogeneity in ccRCC patients and to more accurately identify which metabolic pathways are altered in vivo . Methods 58 tumor and healthy tissues biopsies were acquired postoperatively from 6 ccRCC patients imaged following injection of hyperpolarized [1- 13 C]pyruvate. MRI parameters were correlated with the metabolomic (146 metabolites) and transcriptomic (2523 metabolic genes) data obtained from these biopsies, split across 34 metabolic pathways. The results were used to generate metabologram projections as a visual representation of these metabolic differences. For each metabolic pathway, we generated a novel metabolic consensus scoring system for the identification of key altered metabolic pathways in ccRCC and their relationship to the imaging parameters. Results We show that the apparent exchange constant between pyruvate and lactate ( k PL ) and the lactate to pyruvate ratio (LP r ) on MRI can be used to measure differential metabolic pathways: they correlated positively with glycolysis and the pentose phosphate pathway, negatively with the TCA cycle, while also correlating with other pathways. Dichotomizing the imaged signal based on high and low k PL measurements was sufficient to discriminate metabolic distinct regions on biopsy and this could be a simple tool to assess metabolism clinically. Furthermore, metabolic heterogeneity increased in regions with a higher k PL and could be used to assess metabolic divergence. Conclusion This work validated the role of HP- 13 C-MRI to measure not only glycolysis, but also a range of other altered metabolic pathways in ccRCC. This could improve tumor stratification and provide novel methods to monitor treatment response. Metabolic imaging can also be used to guide biopsy acquisition based on metabolic alterations, and therefore could improve tumour characterization. Clear cell renal cell carcinoma metabolic heterogeneity metabolic imaging hyperpolarized carbon-13 MRI lactate dehydrogenase multiomic analysis novel scoring algorithm multiscale. Full Text Additional Declarations No competing interests reported. Supplementary Files supplementaltable1imagingparametersinindividualbiopsies.xlsx supplementaltable2RNAseqnormalizedgenecountsxlsx.xlsx supplementaltable3metaboliteconcentrationsinindividualbiopsies.xlsx supplementaltable4spearmanandpearsoncorrelationofmetabolicgenesandmetabolicimagingparameters.xlsx supplementaltable5metabolicgenescorrelationparameterssortedbypathway.xlsx supplementaltable6spearmanandpearsoncorrelationofmetabolitesandmetabolicimagingparameters.xlsx supplementaltable7Metabolitecorrelationparameterssortedbypathway.xlsx Supplementalfiguresandlegends.docx graphicalabstract.png Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: Revision requested 16 Feb, 2026 Reviews received at journal 14 Feb, 2026 Reviews received at journal 13 Feb, 2026 Reviewers agreed at journal 01 Feb, 2026 Reviewers agreed at journal 30 Jan, 2026 Reviewers invited by journal 27 Jan, 2026 Editor assigned by journal 31 Oct, 2025 Submission checks completed at journal 31 Oct, 2025 First submitted to journal 30 Oct, 2025 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. 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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-7989219","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":581215874,"identity":"227237b2-82b1-4c75-86fe-eb99a0e92367","order_by":0,"name":"João André Gonçalves Duarte","email":"","orcid":"","institution":"University of Cambridge","correspondingAuthor":false,"prefix":"","firstName":"João","middleName":"André Gonçalves","lastName":"Duarte","suffix":""},{"id":581215875,"identity":"548ca317-b635-4ee6-b8bf-a5c5afefe3b8","order_by":1,"name":"Ines Horvat-Menih","email":"","orcid":"","institution":"University of 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MRI\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-medicine","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bmed","sideBox":"Learn more about [BMC Medicine](http://bmcmedicine.biomedcentral.com/)","snPcode":"12916","submissionUrl":"https://submission.nature.com/new-submission/12916/3","title":"BMC Medicine","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Clear cell renal cell carcinoma, metabolic heterogeneity, metabolic imaging, hyperpolarized carbon-13 MRI, lactate dehydrogenase, multiomic analysis, novel scoring algorithm, multiscale.","lastPublishedDoi":"10.21203/rs.3.rs-7989219/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7989219/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eClear cell renal cell carcinoma (ccRCC) is a highly heterogeneous cancer requiring a large number of biopsies to correctly characterize the tumor. Multiple biopsies are rarely feasible in the clinical setting, and therefore imaging methods offer the potential to evaluate the whole tumor non-invasively. For example, metabolic imaging offers the potential to probe the altered metabolism and metabolic heterogeneity that is characteristic of ccRCC. In this study we have explored and validated the use of hyperpolarized carbon-13 MRI (HP-\u003csup\u003e13\u003c/sup\u003eC-MRI) as a non-invasive clinical tool to probe metabolic heterogeneity in ccRCC patients and to more accurately identify which metabolic pathways are altered \u003cem\u003ein vivo\u003c/em\u003e.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003e58 tumor and healthy tissues biopsies were acquired postoperatively from 6 ccRCC patients imaged following injection of hyperpolarized [1-\u003csup\u003e13\u003c/sup\u003eC]pyruvate. MRI parameters were correlated with the metabolomic (146 metabolites) and transcriptomic (2523 metabolic genes) data obtained from these biopsies, split across 34 metabolic pathways. The results were used to generate metabologram projections as a visual representation of these metabolic differences. For each metabolic pathway, we generated a novel metabolic consensus scoring system for the identification of key altered metabolic pathways in ccRCC and their relationship to the imaging parameters.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eWe show that the apparent exchange constant between pyruvate and lactate (\u003cem\u003ek\u003c/em\u003e\u003csub\u003ePL\u003c/sub\u003e) and the lactate to pyruvate ratio (LP\u003csub\u003er\u003c/sub\u003e) on MRI can be used to measure differential metabolic pathways: they correlated positively with glycolysis and the pentose phosphate pathway, negatively with the TCA cycle, while also correlating with other pathways. Dichotomizing the imaged signal based on high and low \u003cem\u003ek\u003c/em\u003e\u003csub\u003ePL\u003c/sub\u003e measurements was sufficient to discriminate metabolic distinct regions on biopsy and this could be a simple tool to assess metabolism clinically. Furthermore, metabolic heterogeneity increased in regions with a higher \u003cem\u003ek\u003c/em\u003e\u003csub\u003ePL\u003c/sub\u003e and could be used to assess metabolic divergence.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eThis work validated the role of HP-\u003csup\u003e13\u003c/sup\u003eC-MRI to measure not only glycolysis, but also a range of other altered metabolic pathways in ccRCC. This could improve tumor stratification and provide novel methods to monitor treatment response. Metabolic imaging can also be used to guide biopsy acquisition based on metabolic alterations, and therefore could improve tumour characterization.\u003c/p\u003e","manuscriptTitle":"Deconvolving metabolic intratumoral heterogeneity in clear cell renal carcinoma with hyperpolarized 13C-pyruvate MRI","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-29 17:21:35","doi":"10.21203/rs.3.rs-7989219/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-02-16T09:41:16+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-14T06:51:08+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-13T14:46:56+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"45862967254074641459803498185205596419","date":"2026-02-01T12:10:29+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"197116911566619313855966072906926180473","date":"2026-01-30T08:34:09+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-01-27T14:20:16+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-10-31T06:23:18+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-10-31T06:04:56+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Medicine","date":"2025-10-30T12:04:09+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-medicine","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bmed","sideBox":"Learn more about [BMC Medicine](http://bmcmedicine.biomedcentral.com/)","snPcode":"12916","submissionUrl":"https://submission.nature.com/new-submission/12916/3","title":"BMC Medicine","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"9e106851-f0d2-47b2-9c5d-1641163e9552","owner":[],"postedDate":"January 29th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[],"tags":[],"updatedAt":"2026-02-16T09:56:24+00:00","versionOfRecord":[],"versionCreatedAt":"2026-01-29 17:21:35","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7989219","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7989219","identity":"rs-7989219","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}