Foxk2 regulates osteoblast differentiation and bone formation through Slc1a4-mediated enhancement of glutamine-dependent energy metabolism

Foxk2 regulates osteoblast differentiation and bone formation through Slc1a4-mediated enhancement of glutamine-dependent energy metabolism | 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 Foxk2 regulates osteoblast differentiation and bone formation through Slc1a4-mediated enhancement of glutamine-dependent energy metabolism Songlin Peng, Chungeng Liu, Junfei Zhao, Jian Liu, Zhenmin Wang, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8690219/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 Foxk2 can regulate cell differentiation, proliferation, metabolism and tumor fate. However, the roles of Foxk2 in bone formation are unknown. Here, we found that global knockout of Foxk2 in mice resulted in decreased bone mass and bone formation. Deletion of Foxk2 in primary pre-osteoblasts suppressed osteoblast differentiation in vitro and conditional knockout of Foxk2 in pre-osteoblasts in mice showed attenuated bone mass and bone formation. Notably, overexpression of Foxk2 in pre-osteoblasts alleviated bone loss in mice caused by ovariectomy. Mechanistically, we identified Slc1a4 as a downstream target of Foxk2 through joint analysis the data of RNA-Seq and CUT&Tag. Knockout of Slc1a4 in pre-osteoblasts in mice resulted in decreased bone mass and bone formation. Overexpression of Slc1a4 in pre-osteoblasts alleviated bone loss and damaged bone formation caused by Foxk2 deletion. Furthermore, metabolomic analyses highlighted the role for Foxk2 in promoting glutamine-dependent energy metabolism mediated by Slc1a4, which is required for osteoblast differentiation. In summary, our study demonstrated that Foxk2 promotes osteoblast differentiation and bone formation through Slc1a4-mediated enhancement of glutamine-dependent energy metabolism, which is also a promising target for preventing osteoporosis. Biological sciences/Physiology/Bone Biological sciences/Stem cells/Stem-cell differentiation Health sciences/Diseases/Endocrine system and metabolic diseases/Metabolic bone disease/Osteoporosis Full Text Additional Declarations There is NO Competing Interest. Supplementary Files Supplementarymaterials.pdf Supplementary materials 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. 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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-8690219","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":597277444,"identity":"d1e3e685-a6d9-4aa3-a031-92897aec094f","order_by":0,"name":"Songlin Peng","email":"data:image/png;base64,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","orcid":"","institution":"Shenzhen People's Hospital(The Second Clinical Medical College, Jinan University; 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However, the roles of Foxk2 in bone formation are unknown. Here, we found that global knockout of Foxk2 in mice resulted in decreased bone mass and bone formation. Deletion of Foxk2 in primary pre-osteoblasts suppressed osteoblast differentiation in vitro and conditional knockout of Foxk2 in pre-osteoblasts in mice showed attenuated bone mass and bone formation. Notably, overexpression of Foxk2 in pre-osteoblasts alleviated bone loss in mice caused by ovariectomy. Mechanistically, we identified Slc1a4 as a downstream target of Foxk2 through joint analysis the data of RNA-Seq and CUT\u0026Tag. Knockout of Slc1a4 in pre-osteoblasts in mice resulted in decreased bone mass and bone formation. Overexpression of Slc1a4 in pre-osteoblasts alleviated bone loss and damaged bone formation caused by Foxk2 deletion. Furthermore, metabolomic analyses highlighted the role for Foxk2 in promoting glutamine-dependent energy metabolism mediated by Slc1a4, which is required for osteoblast differentiation. In summary, our study demonstrated that Foxk2 promotes osteoblast differentiation and bone formation through Slc1a4-mediated enhancement of glutamine-dependent energy metabolism, which is also a promising target for preventing osteoporosis.","manuscriptTitle":"Foxk2 regulates osteoblast differentiation and bone formation through Slc1a4-mediated enhancement of glutamine-dependent energy metabolism","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-03-03 21:27:05","doi":"10.21203/rs.3.rs-8690219/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":"704c0d9f-217f-4072-8a04-8041af62bed0","owner":[],"postedDate":"March 3rd, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":63556133,"name":"Biological sciences/Physiology/Bone"},{"id":63556134,"name":"Biological sciences/Stem cells/Stem-cell differentiation"},{"id":63556135,"name":"Health sciences/Diseases/Endocrine system and metabolic diseases/Metabolic bone disease/Osteoporosis"}],"tags":[],"updatedAt":"2026-03-31T09:29:20+00:00","versionOfRecord":[],"versionCreatedAt":"2026-03-03 21:27:05","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8690219","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8690219","identity":"rs-8690219","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}