Dusp15 Dephosphorylates mtHsp70 at Thr116 to Prevent Its Proteasomal Degradation and Maintain Mitochondrial Unfolded Protein Response in Diabetic Cardiomyopathy

Dusp15 Dephosphorylates mtHsp70 at Thr116 to Prevent Its Proteasomal Degradation and Maintain Mitochondrial Unfolded Protein Response in Diabetic Cardiomyopathy | 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 Dusp15 Dephosphorylates mtHsp70 at Thr116 to Prevent Its Proteasomal Degradation and Maintain Mitochondrial Unfolded Protein Response in Diabetic Cardiomyopathy xing chang, Rongjun Zou, Jinlin Hu, Miao Zhang, Hongshuo Shi, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6428877/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 Diabetic cardiomyopathy (DCM) is driven by mitochondrial dysfunction and impaired proteostasis. This study investigates Dusp15’s role in regulating the mitochondrial unfolded protein response (mito-UPR) via its interaction with mtHsp70 and explores its therapeutic potential in DCM. Transcriptomic and proteomic analyses identified Dusp15 as a critical regulator in DCM. Overexpression of Dusp15 preserved cardiac function, reduced fibrosis and inflammation, and stabilized mitochondrial function while Dusp15 deficiency exacerbated these pathologies. Mechanistically, Dusp15 interacted with the N-terminal binding domain (NBD) of mtHsp70 via its phosphatase domain, targeting Thr116 within the TFY motif for dephosphorylation. This prevented mtHsp70 proteasomal degradation, maintained mito-UPR activity, mitochondrial membrane potential, and redox balance. mtHsp70 phosphorylation-deficient mice ( mtHsp70T116A ) mice resisted hyperglycemia-induced myocardial dysfunction, fibrosis, and inflammation, while phosphorylation-mimetic T116D mutants negated Dusp15-mediated cardioprotection. Therapeutically, a drug screen integrating RNA sequencing and pathway scoring identified dapagliflozin (DAPA) as a cardioprotective candidate. DAPA upregulated Dusp15, stabilized mtHsp70, and improved cardiac outcomes, effects absent in Dusp15 Cko mice. Inhibition of mito-UPR negated Dusp15 and DAPA benefits. Dusp15 preserves mitochondrial integrity and cardiac function in DCM by dephosphorylating mtHsp70 at Thr116. Targeting the Dusp15/mtHsp70 axis, including with DAPA, offers a promising strategy for DCM treatment. Health sciences/Biomarkers Health sciences/Diseases/Cardiovascular diseases Biological sciences/Cell biology Diabetic cardiomyopathy Dusp15 mtHsp70 mitochondrial unfolded protein response Full Text Additional Declarations There is NO Competing Interest. Supplementary Files Graphabstract.jpg Dusp15 Dephosphorylates mtHsp70 at Thr116 to Prevent Its Proteasomal Degradation and Maintain Mitochondrial Unfolded Protein Response in Diabetic Cardiomyopathy Supplementalfiguresandtables.pdf Dataset 1 49Highlights.docx Dataset 1 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. 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Cardiomyopathy","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"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":"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":"Diabetic cardiomyopathy, Dusp15, mtHsp70, mitochondrial unfolded protein response","lastPublishedDoi":"10.21203/rs.3.rs-6428877/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6428877/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eDiabetic cardiomyopathy (DCM) is driven by mitochondrial dysfunction and impaired proteostasis. This study investigates Dusp15\u0026rsquo;s role in regulating the mitochondrial unfolded protein response (mito-UPR) via its interaction with mtHsp70 and explores its therapeutic potential in DCM. Transcriptomic and proteomic analyses identified Dusp15 as a critical regulator in DCM. Overexpression of \u003cem\u003eDusp15\u003c/em\u003e preserved cardiac function, reduced fibrosis and inflammation, and stabilized mitochondrial function while \u003cem\u003eDusp15\u003c/em\u003e deficiency exacerbated these pathologies. Mechanistically, Dusp15 interacted with the N-terminal binding domain (NBD) of mtHsp70 via its phosphatase domain, targeting Thr116 within the TFY motif for dephosphorylation. This prevented mtHsp70 proteasomal degradation, maintained mito-UPR activity, mitochondrial membrane potential, and redox balance. mtHsp70 phosphorylation-deficient mice (\u003cem\u003emtHsp70T116A\u003c/em\u003e) mice resisted hyperglycemia-induced myocardial dysfunction, fibrosis, and inflammation, while phosphorylation-mimetic T116D mutants negated Dusp15-mediated cardioprotection. Therapeutically, a drug screen integrating RNA sequencing and pathway scoring identified dapagliflozin (DAPA) as a cardioprotective candidate. DAPA upregulated Dusp15, stabilized mtHsp70, and improved cardiac outcomes, effects absent in \u003cem\u003eDusp15\u003c/em\u003e\u003csup\u003e\u003cem\u003eCko\u003c/em\u003e\u003c/sup\u003e mice. Inhibition of mito-UPR negated Dusp15 and DAPA benefits. Dusp15 preserves mitochondrial integrity and cardiac function in DCM by dephosphorylating mtHsp70 at Thr116. Targeting the Dusp15/mtHsp70 axis, including with DAPA, offers a promising strategy for DCM treatment.\u003c/p\u003e","manuscriptTitle":"Dusp15 Dephosphorylates mtHsp70 at Thr116 to Prevent Its Proteasomal Degradation and Maintain Mitochondrial Unfolded Protein Response in Diabetic Cardiomyopathy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-21 15:35:54","doi":"10.21203/rs.3.rs-6428877/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":"b60240cf-be95-4712-9974-7232f9ec56bd","owner":[],"postedDate":"May 21st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":48778410,"name":"Health sciences/Biomarkers"},{"id":48778411,"name":"Health sciences/Diseases/Cardiovascular diseases"},{"id":48778412,"name":"Biological sciences/Cell biology"}],"tags":[],"updatedAt":"2025-07-16T05:15:13+00:00","versionOfRecord":[],"versionCreatedAt":"2025-05-21 15:35:54","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6428877","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6428877","identity":"rs-6428877","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}