Metabolic Adaptations to Caloric Restriction: Time- and Group-Dependent Metabolomic Signatures from the CALERIE™ Trial

Metabolic Adaptations to Caloric Restriction: Time- and Group-Dependent Metabolomic Signatures from the CALERIE™ Trial | 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 Metabolic Adaptations to Caloric Restriction: Time- and Group-Dependent Metabolomic Signatures from the CALERIE™ Trial Melissa C. Orenduff, Emily K. Woolf, Ruiqi Zhang, Daniel W. Belsky, and 9 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9237764/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract Background : Caloric restriction (CR) improves markers of biological aging, yet long-term effects on the human metabolome remain unclear. Objective: This study examined the effects of CR (2 years) in healthy adults without obesity on circulating metabolites linked to aging and metabolic adaptations. Methods: Untargeted metabolomics was performed using fasted plasma samples collected at baseline, 12, and 24 months (BL, 12M, 24M) from CALERIE™ participants randomized to CR or ad libitum (AL) control. A total of 864 known metabolites were identified and grouped into nine biologically coherent super pathways to support pathway‑level interpretation (amino acid, peptide, carbohydrate, energy, lipid, nucleotide, cofactors and vitamins, xenobiotics, and partially characterized molecules). Principal component analysis (PCA) summarized metabolite variation, and linear mixed models assessed intervention effects on each PC in group-by-time interactions. Results: Three principal components showed significant group‑by‑time interactions: PC2 (carbohydrate), PC5 (partially characterized molecules), and PC4 (lipid). Carbohydrate (PC2) and partially characterized metabolites (PC5) decreased from baseline to 12M in both groups; from 12M to 24M, levels stabilized in CR but increased in AL for PC2, while PC5 continued to decline in AL and increased in CR. Lipid metabolites (PC4) decreased in CR and increased in AL at 12M, with the pattern reversing from 12M to 24M. Key contributors included malto‑saccharides and related carbohydrate intermediates for PC2, glutamine degradants and lactone sulfates for PC5, and sphingolipids for PC4. Conclusion: Calorie restriction produced distinct, time‑dependent shifts in carbohydrate and lipid metabolism, with early reductions during the weight‑loss phase followed by stabilization or compensatory responses during weight maintenance. These dynamic metabolic changes may relate to inflammation‑linked mechanisms. Further work is needed to distinguish CR‑specific adaptations from dietary influences and to clarify the functional significance of these metabolic responses for aging and long‑term metabolic health. CALERIE™ Caloric Restriction Metabolomics principal component analysis (PCA) Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementalTables123CALERIEmetabolomicsmanuscript.docx Cite Share Download PDF Status: Under Review Version 1 posted Reviewers agreed at journal 25 Apr, 2026 Reviewers agreed at journal 23 Apr, 2026 Reviews received at journal 20 Apr, 2026 Reviews received at journal 16 Apr, 2026 Reviewers agreed at journal 09 Apr, 2026 Reviewers agreed at journal 27 Mar, 2026 Reviewers invited by journal 27 Mar, 2026 Editor assigned by journal 27 Mar, 2026 Submission checks completed at journal 27 Mar, 2026 First submitted to journal 26 Mar, 2026 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-9237764","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":613162687,"identity":"3d5b0d54-9642-4909-a373-bff2c0aff94b","order_by":0,"name":"Melissa C. 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A total of 864 known metabolites were identified and grouped into nine biologically coherent super pathways to support pathway‑level interpretation (amino acid, peptide, carbohydrate, energy, lipid, nucleotide, cofactors and vitamins, xenobiotics, and partially characterized molecules). Principal component analysis (PCA) summarized metabolite variation, and linear mixed models assessed intervention effects on each PC in group-by-time interactions.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eThree principal components showed significant group‑by‑time interactions: PC2 (carbohydrate), PC5 (partially characterized molecules), and PC4 (lipid). Carbohydrate (PC2) and partially characterized metabolites (PC5) decreased from baseline to 12M in both groups; from 12M to 24M, levels stabilized in CR but increased in AL for PC2, while PC5 continued to decline in AL and increased in CR. Lipid metabolites (PC4) decreased in CR and increased in AL at 12M, with the pattern reversing from 12M to 24M. Key contributors included malto‑saccharides and related carbohydrate intermediates for PC2, glutamine degradants and lactone sulfates for PC5, and sphingolipids for PC4.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCalorie restriction produced distinct, time‑dependent shifts in carbohydrate and lipid metabolism, with early reductions during the weight‑loss phase followed by stabilization or compensatory responses during weight maintenance. These dynamic metabolic changes may relate to inflammation‑linked mechanisms. Further work is needed to distinguish CR‑specific adaptations from dietary influences and to clarify the functional significance of these metabolic responses for aging and long‑term metabolic health.\u003c/p\u003e","manuscriptTitle":"Metabolic Adaptations to Caloric Restriction: Time- and Group-Dependent Metabolomic Signatures from the CALERIE™ Trial","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-03-30 08:31:08","doi":"10.21203/rs.3.rs-9237764/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"239202517855538783814612522454201414248","date":"2026-04-25T11:44:30+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"287067663516105648959679012031623655430","date":"2026-04-24T00:21:56+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-20T12:46:37+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-16T21:00:48+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"303820472136429307365638224195464241507","date":"2026-04-09T16:17:54+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"106871664936689160069195746829787220304","date":"2026-03-27T08:54:24+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-03-27T08:40:41+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-03-27T06:39:45+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-03-27T06:39:10+00:00","index":"","fulltext":""},{"type":"submitted","content":"Metabolomics","date":"2026-03-26T19:54:47+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"metabolomics","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mebo","sideBox":"Learn more about [Metabolomics](http://link.springer.com/journal/11306)","snPcode":"11306","submissionUrl":"https://submission.nature.com/new-submission/11306/3","title":"Metabolomics","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"6fc0fe2e-ce4b-45d9-b323-746d445599ff","owner":[],"postedDate":"March 30th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-03-30T08:31:08+00:00","versionOfRecord":[],"versionCreatedAt":"2026-03-30 08:31:08","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9237764","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9237764","identity":"rs-9237764","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}