Regulators of health and lifespan extension in genetically diverse mice on dietary restriction

Regulators of health and lifespan extension in genetically diverse mice on dietary restriction | 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 Biological Sciences - Article Regulators of health and lifespan extension in genetically diverse mice on dietary restriction Gary Churchill, Andrea Di Francesco, Andrew Deighan, Lev Litichevskiy, and 18 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3678113/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 09 Oct, 2024 Read the published version in Nature → Version 1 posted You are reading this latest preprint version Abstract Caloric restriction (CR) delays aging and extends healthy lifespan in multiple species. Alternative forms of dietary restriction (DR) such as intermittent fasting (IF) have drawn significant interest as a more sustainable regimen, but the landscape of longevity-promoting dietary interventions remains largely unexplored. Identifying the most robust, efficacious, and experimentally tractable modes of DR is key to better understanding and implementing effective longevity interventions for human healthspan. To that end, we have performed an extensive assessment of DR interventions, investigating the effects of graded levels of CR (20% and 40%) and IF (1 day and 2 days of fasting per week) on the health and survival of 960 genetically diverse female mice. All interventions extended lifespan, although only CR significantly reduced the mortality doubling time. Notably, IF did not extend lifespan in mice with high pre-intervention bodyweight. We carried out extensive phenotyping to determine the health effects of long-term DR and to better understand the mechanisms driving within-diet heterogeneity in lifespan. The top within-diet predictor of lifespan was the ability of mice to maintain bodyweight through periods of handling, an indicator of stress resilience. Additional predictors of long lifespan include specific changes in immune cells, red blood cell distribution width (RDW), and retention of adiposity in late life. We found that lifespan is heritable (h 2 = 0.24), and that genetic background has a larger influence on lifespan than dietary interventions. We identified a significant association for lifespan and RDW on chromosome 18 that explained 4.3% of the diet-adjusted variation in lifespan. Diet-induced changes on metabolic traits, although beneficial, were relatively poor predictors of lifespan, arguing against the long-standing notion that DR works by counteracting the negative effects of obesity. These findings indicate that improving health and extending lifespan are not synonymous and that metabolic parameters may be inappropriate endpoints for evaluating aging interventions in preclinical models and clinical trials. Biological sciences/Physiology/Ageing Biological sciences/Genetics Full Text Additional Declarations Yes there is potential Competing Interest. ADF, ZC, KMW, AR, GVP, MM, and FH, RLC are employees of Calico Life Sciences LLC. Supplementary Files SupplementalTable4DietAgeBW.csv Dataset 4 SupplementalTable5SurvCor.csv Dataset 5 SupplementalTable6NetworkDetails.xlsx Dataset 6 SupplementalTable7FACSNames.xlsx Dataset 7 Cite Share Download PDF Status: Published Journal Publication published 09 Oct, 2024 Read the published version in Nature → 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-3678113","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Biological Sciences - Article","associatedPublications":[],"authors":[{"id":265429256,"identity":"d8b8c500-f609-4b75-9e07-bd80e8c49ccf","order_by":0,"name":"Gary Churchill","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA+0lEQVRIiWNgGAWjYBAC/gYQWXMASPAwHHhQYMNMUIsESDHDMaiWBIM0wloMQARjE0QLQ4LBYcIOM2A/+0yCseFO4vb2sweBtpxn55+RwPi44hceLTzpZkAtzxLnnMlLAGq5zSxx5gCz4dk+3FoMD6SxAbUcTpwhwWMA1sJwvIFNsrEHjy3nn6FoOccsf5iBgJYbIFua4FoOMBuAbGn4gVuLxI1nzBYJxw4bz+DJAWlJZjY8c7DZsLEBtxb+/jTGGx9qDsvOYD9j/OFDhV2y3I3kgw8b/uDWAgQsEglIvGRgNDUwMLbh1cL8AZlnB6Hw2zIKRsEoGAUjCwAAkVJYBj+Uso8AAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0001-9190-9284","institution":"Jackson Laboratories","correspondingAuthor":true,"prefix":"","firstName":"Gary","middleName":"","lastName":"Churchill","suffix":""},{"id":265429257,"identity":"28c75108-1d77-4c4b-b6eb-314f093db953","order_by":1,"name":"Andrea Di Francesco","email":"","orcid":"","institution":"Calico Life Sciences LL","correspondingAuthor":false,"prefix":"","firstName":"Andrea","middleName":"Di","lastName":"Francesco","suffix":""},{"id":265429258,"identity":"732c3684-4619-4327-bd4c-42c7d8949ade","order_by":2,"name":"Andrew Deighan","email":"","orcid":"","institution":"The Jackson Laboratory","correspondingAuthor":false,"prefix":"","firstName":"Andrew","middleName":"","lastName":"Deighan","suffix":""},{"id":265429259,"identity":"4ccd7ae4-9ee4-4eea-93bc-62913f5d8451","order_by":3,"name":"Lev Litichevskiy","email":"","orcid":"https://orcid.org/0000-0002-0510-4509","institution":"University of Pennsylvania","correspondingAuthor":false,"prefix":"","firstName":"Lev","middleName":"","lastName":"Litichevskiy","suffix":""},{"id":265429260,"identity":"ec60fe42-df33-411f-ab4d-3c72896ed701","order_by":4,"name":"Zhenghao Chen","email":"","orcid":"","institution":"Calico Life Sciences LLC","correspondingAuthor":false,"prefix":"","firstName":"Zhenghao","middleName":"","lastName":"Chen","suffix":""},{"id":265429261,"identity":"b8c47eb5-f9f2-41fd-bc10-42fd44b68265","order_by":5,"name":"Alison Luciano","email":"","orcid":"","institution":"The Jackson Laboratory","correspondingAuthor":false,"prefix":"","firstName":"Alison","middleName":"","lastName":"Luciano","suffix":""},{"id":265429262,"identity":"d89d3c2c-cedb-4d5b-820e-a82076e2595c","order_by":6,"name":"Laura Robinson","email":"","orcid":"","institution":"The Jackson Laboratory","correspondingAuthor":false,"prefix":"","firstName":"Laura","middleName":"","lastName":"Robinson","suffix":""},{"id":265429263,"identity":"836d811c-361d-497d-b85f-75865b268777","order_by":7,"name":"Gaven Garland","email":"","orcid":"","institution":"The Jackson Laboratory","correspondingAuthor":false,"prefix":"","firstName":"Gaven","middleName":"","lastName":"Garland","suffix":""},{"id":265429264,"identity":"49e91ad4-0433-46a8-bd29-05146b87734b","order_by":8,"name":"Hannah Donato","email":"","orcid":"","institution":"The Jackson Laboratory","correspondingAuthor":false,"prefix":"","firstName":"Hannah","middleName":"","lastName":"Donato","suffix":""},{"id":265429265,"identity":"690b3fc2-f2a4-431f-84e1-d333ec8b2e99","order_by":9,"name":"Will Schott","email":"","orcid":"","institution":"The Jackson Laboratory","correspondingAuthor":false,"prefix":"","firstName":"Will","middleName":"","lastName":"Schott","suffix":""},{"id":265429266,"identity":"b0671879-52bf-40f3-9922-115a2372bc45","order_by":10,"name":"Kevin Wright","email":"","orcid":"","institution":"Calico","correspondingAuthor":false,"prefix":"","firstName":"Kevin","middleName":"","lastName":"Wright","suffix":""},{"id":265429267,"identity":"31cfb99d-9c94-4996-a90b-1f156f64e71d","order_by":11,"name":"Anil Raj","email":"","orcid":"","institution":"Calico","correspondingAuthor":false,"prefix":"","firstName":"Anil","middleName":"","lastName":"Raj","suffix":""},{"id":265429268,"identity":"36dc4674-eb71-4e5b-b33c-491d3cec17d7","order_by":12,"name":"G.V. Prateek","email":"","orcid":"","institution":"Calico Life Sciences LLC","correspondingAuthor":false,"prefix":"","firstName":"G.V.","middleName":"","lastName":"Prateek","suffix":""},{"id":265429269,"identity":"e7c64c49-ec61-43a7-aa07-e2babc1bfbac","order_by":13,"name":"Martin Mullis","email":"","orcid":"https://orcid.org/0000-0002-5134-7528","institution":"Calico Life Sciences LLC","correspondingAuthor":false,"prefix":"","firstName":"Martin","middleName":"","lastName":"Mullis","suffix":""},{"id":265429270,"identity":"c631d4f3-98ee-4293-9862-0b7a8f58224f","order_by":14,"name":"Warren Hill","email":"","orcid":"","institution":"Beth Israel Deaconess Medical Center","correspondingAuthor":false,"prefix":"","firstName":"Warren","middleName":"","lastName":"Hill","suffix":""},{"id":265429271,"identity":"d26c08be-ccb3-4123-979a-aa62e1da78c7","order_by":15,"name":"Mark Zeidel","email":"","orcid":"","institution":"Beth Israel Deaconess Medical Center","correspondingAuthor":false,"prefix":"","firstName":"Mark","middleName":"","lastName":"Zeidel","suffix":""},{"id":265429272,"identity":"13dc1681-0a15-43cc-895b-e245552c9295","order_by":16,"name":"Luanne Peters","email":"","orcid":"","institution":"The Jackson Laboratory, Bar Harbor, ME 04609, USA","correspondingAuthor":false,"prefix":"","firstName":"Luanne","middleName":"","lastName":"Peters","suffix":""},{"id":265429273,"identity":"b4b4eac4-70d7-45d5-a0fe-f5213553586e","order_by":17,"name":"Fiona Harding","email":"","orcid":"","institution":"Calico Life Sciences LLC","correspondingAuthor":false,"prefix":"","firstName":"Fiona","middleName":"","lastName":"Harding","suffix":""},{"id":265429274,"identity":"97fd50ec-7492-408e-8493-a0a10bf1060d","order_by":18,"name":"David Botstein","email":"","orcid":"","institution":"Calico Life Sciences LLC","correspondingAuthor":false,"prefix":"","firstName":"David","middleName":"","lastName":"Botstein","suffix":""},{"id":265429275,"identity":"7e3a0f27-a971-4633-b287-cb9f0468613a","order_by":19,"name":"Ron Korstanje","email":"","orcid":"https://orcid.org/0000-0002-2808-1610","institution":"The Jackson Laboratory","correspondingAuthor":false,"prefix":"","firstName":"Ron","middleName":"","lastName":"Korstanje","suffix":""},{"id":265429276,"identity":"95305898-d9de-4bde-89d6-fba03c8bcdfd","order_by":20,"name":"Christoph Thaiss","email":"","orcid":"https://orcid.org/0000-0001-8226-7718","institution":"University of Pennsylvania","correspondingAuthor":false,"prefix":"","firstName":"Christoph","middleName":"","lastName":"Thaiss","suffix":""},{"id":265429277,"identity":"f1403dce-7b6b-46f8-b3dc-85b9b5b174c3","order_by":21,"name":"Adam Freund","email":"","orcid":"https://orcid.org/0000-0002-7956-5332","institution":"Calico Life Sciences LLC","correspondingAuthor":false,"prefix":"","firstName":"Adam","middleName":"","lastName":"Freund","suffix":""}],"badges":[],"createdAt":"2023-11-28 19:10:31","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3678113/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3678113/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41586-024-08026-3","type":"published","date":"2024-10-09T04:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":66304092,"identity":"ec5e6a1b-a385-46e7-b901-cb21490037af","added_by":"auto","created_at":"2024-10-10 07:08:17","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":6872257,"visible":true,"origin":"","legend":"Article File","description":"","filename":"DRiDOanchormanuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3678113/v1_covered_6c0f128a-4016-41b4-963d-c1483b19c1e2.pdf"},{"id":49285232,"identity":"6d420da5-8312-4eb3-995f-b0989b5b3982","added_by":"auto","created_at":"2024-01-08 01:35:38","extension":"csv","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":32304,"visible":true,"origin":"","legend":"Dataset 4","description":"","filename":"SupplementalTable4DietAgeBW.csv","url":"https://assets-eu.researchsquare.com/files/rs-3678113/v1/8bb2d627bf418a79b9f9ce3f.csv"},{"id":49285429,"identity":"41eff0f9-728c-496e-b15e-00b996340aa8","added_by":"auto","created_at":"2024-01-08 01:43:38","extension":"csv","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":157748,"visible":true,"origin":"","legend":"\u003cp\u003eDataset 5\u003c/p\u003e","description":"","filename":"SupplementalTable5SurvCor.csv","url":"https://assets-eu.researchsquare.com/files/rs-3678113/v1/b987252fca956f5be1a4ac55.csv"},{"id":49285235,"identity":"9c94aba1-9993-4f52-90e0-d93e93334edc","added_by":"auto","created_at":"2024-01-08 01:35:38","extension":"xlsx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":17201,"visible":true,"origin":"","legend":"\u003cp\u003eDataset 6\u003c/p\u003e","description":"","filename":"SupplementalTable6NetworkDetails.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-3678113/v1/a12f2dcf3dc2e5b8a2f7ede7.xlsx"},{"id":49285728,"identity":"a84d8b3e-f932-4daa-b2c6-b996b7356718","added_by":"auto","created_at":"2024-01-08 01:51:38","extension":"xlsx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":14045,"visible":true,"origin":"","legend":"\u003cp\u003eDataset 7\u003c/p\u003e","description":"","filename":"SupplementalTable7FACSNames.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-3678113/v1/12219b42959ee237073279cf.xlsx"}],"financialInterests":"\u003cb\u003eYes\u003c/b\u003e there is potential Competing Interest.\nADF, ZC, KMW, AR, GVP, MM, and FH, RLC are employees of Calico Life Sciences LLC.","formattedTitle":"Regulators of health and lifespan extension in genetically diverse mice on dietary restriction","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-3678113/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3678113/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eCaloric restriction (CR) delays aging and extends healthy lifespan in multiple species. Alternative forms of dietary restriction (DR) such as intermittent fasting (IF) have drawn significant interest as a more sustainable regimen, but the landscape of longevity-promoting dietary interventions remains largely unexplored. Identifying the most robust, efficacious, and experimentally tractable modes of DR is key to better understanding and implementing effective longevity interventions for human healthspan. To that end, we have performed an extensive assessment of DR interventions, investigating the effects of graded levels of CR (20% and 40%) and IF (1 day and 2 days of fasting per week) on the health and survival of 960 genetically diverse female mice. All interventions extended lifespan, although only CR significantly reduced the mortality doubling time. Notably, IF did not extend lifespan in mice with high pre-intervention bodyweight. We carried out extensive phenotyping to determine the health effects of long-term DR and to better understand the mechanisms driving within-diet heterogeneity in lifespan. The top within-diet predictor of lifespan was the ability of mice to maintain bodyweight through periods of handling, an indicator of stress resilience. Additional predictors of long lifespan include specific changes in immune cells, red blood cell distribution width (RDW), and retention of adiposity in late life. We found that lifespan is heritable (h\u003csup\u003e2\u003c/sup\u003e = 0.24), and that genetic background has a larger influence on lifespan than dietary interventions. We identified a significant association for lifespan and RDW on chromosome 18 that explained 4.3% of the diet-adjusted variation in lifespan. Diet-induced changes on metabolic traits, although beneficial, were relatively poor predictors of lifespan, arguing against the long-standing notion that DR works by counteracting the negative effects of obesity. These findings indicate that improving health and extending lifespan are not synonymous and that metabolic parameters may be inappropriate endpoints for evaluating aging interventions in preclinical models and clinical trials.\u003c/p\u003e","manuscriptTitle":"Regulators of health and lifespan extension in genetically diverse mice on dietary restriction","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-01-08 01:35:34","doi":"10.21203/rs.3.rs-3678113/v1","editorialEvents":[],"status":"published","journal":{"display":false,"email":"[email protected]","identity":"nature","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"nature","sideBox":"Learn more about [Nature](http://www.nature.com/nature/)","snPcode":"","submissionUrl":"","title":"Nature","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"ca967533-86dc-4560-b663-99d499f3890a","owner":[],"postedDate":"January 8th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":27977924,"name":"Biological sciences/Physiology/Ageing"},{"id":27977925,"name":"Biological sciences/Genetics"}],"tags":[],"updatedAt":"2024-10-10T07:08:04+00:00","versionOfRecord":{"articleIdentity":"rs-3678113","link":"https://doi.org/10.1038/s41586-024-08026-3","journal":{"identity":"nature","isVorOnly":false,"title":"Nature"},"publishedOn":"2024-10-09 04:00:00","publishedOnDateReadable":"October 9th, 2024"},"versionCreatedAt":"2024-01-08 01:35:34","video":"","vorDoi":"10.1038/s41586-024-08026-3","vorDoiUrl":"https://doi.org/10.1038/s41586-024-08026-3","workflowStages":[]},"version":"v1","identity":"rs-3678113","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3678113","identity":"rs-3678113","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}