Rejuvenation driven reprograming in T lymphocytes

Rejuvenation driven reprograming in T lymphocytes | 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 Letter Rejuvenation driven reprograming in T lymphocytes Alessio Lanna, Clara D'Ambra, Manuel Delpero, Melania Capitani, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5180379/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 Rejuvenation restores cellular function lost with age, but it is not known to confer properties the cell lacked in its younger past. We found that Disruptors of sestrin-MAPK interactions (DOS) reprogramed senescent T cells into new stem like T clones with different T cell receptors (TCRs). DOS targeted sestrin-MAPK complexes (sMAC) to ubiquitin-dependent proteasomal degradation, resulting in long-term sestrin transcriptional inhibition, increased T cell fitness, and generation of long-lived stem like memory clones. Strikingly, DOS reactivated juvenile related Rag re-expression, leading to antigen-specific TCR rearrangements in formerly senescent cells. As such, rejuvenated T cells, with stem features and new TCRs, combated multiple lethal challenges never previously encountered in their lifetime, with no need for vaccination. T cell rejuvenation confers new properties whilst preserving cellular state, which unravels an unknown rejuvenating principle in biological systems. Biological sciences/Immunology/Signal transduction Biological sciences/Immunology/Lymphocytes/T cells Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Full Text Additional Declarations Yes there is potential Competing Interest. A.L. is a shareholder to Sentcell ltd and the sole inventor of the DOS pharmaceutics where Sentcell ltd figures as the Applicant (PCT/IT2021/000059). M.K. serves as a scientific advisor to Sentcell ltd. A.L. M.C and F.R. are supported by Sentcell ltd. Supplementary Files NCBED1.pdf NCBED2.pdf NCBED3.pdf NCBED4.pdf NCBED5.pdf NCBED6.pdf NCBED7.pdf NCBED8.pdf NCBED9.pdf NCBED10.pdf NCBED11.pdf NCBED12.pdf NCBED13.pdf NCBED14.pdf NCBED15.pdf NCBED16.pdf 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. 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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-5180379","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Letter","associatedPublications":[],"authors":[{"id":363311507,"identity":"9defb24b-eeac-4222-9f6f-e17e005d17dc","order_by":0,"name":"Alessio Lanna","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0003-2215-3748","institution":"Sentcell","correspondingAuthor":true,"prefix":"","firstName":"Alessio","middleName":"","lastName":"Lanna","suffix":""},{"id":363311508,"identity":"f5105d88-3df8-4044-88ff-a229da937fdf","order_by":1,"name":"Clara D'Ambra","email":"","orcid":"","institution":"Sentcell","correspondingAuthor":false,"prefix":"","firstName":"Clara","middleName":"","lastName":"D'Ambra","suffix":""},{"id":363311509,"identity":"d6b7ea3a-83e0-412c-b962-5f05732c913c","order_by":2,"name":"Manuel Delpero","email":"","orcid":"","institution":"MGI","correspondingAuthor":false,"prefix":"","firstName":"Manuel","middleName":"","lastName":"Delpero","suffix":""},{"id":363311510,"identity":"d2f85379-e3ee-4393-a8b6-77e9427f58e5","order_by":3,"name":"Melania Capitani","email":"","orcid":"","institution":"Sentcell","correspondingAuthor":false,"prefix":"","firstName":"Melania","middleName":"","lastName":"Capitani","suffix":""},{"id":363311511,"identity":"90f77323-5ca1-43b4-9ec0-5403b0f1c71a","order_by":4,"name":"Luisa Chocarro","email":"","orcid":"","institution":"Pamplona University","correspondingAuthor":false,"prefix":"","firstName":"Luisa","middleName":"","lastName":"Chocarro","suffix":""},{"id":363311512,"identity":"300ce09e-d57d-4c93-8edf-b71ab962105f","order_by":5,"name":"Federica Rinaldi","email":"","orcid":"","institution":"Sentcell","correspondingAuthor":false,"prefix":"","firstName":"Federica","middleName":"","lastName":"Rinaldi","suffix":""},{"id":363311513,"identity":"842bd7d5-94fc-49d3-a942-3210c743b1ce","order_by":6,"name":"Michael Karin","email":"","orcid":"","institution":"UCSD","correspondingAuthor":false,"prefix":"","firstName":"Michael","middleName":"","lastName":"Karin","suffix":""}],"badges":[],"createdAt":"2024-09-30 11:21:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5180379/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5180379/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":66297977,"identity":"4cce0e8c-7dc1-40e6-9446-91e2f1147f62","added_by":"auto","created_at":"2024-10-10 04:12:14","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1081357,"visible":true,"origin":"","legend":"\u003cp\u003eDiscovery of DOS a, Molecular structure of the lead compound DOS46L (hereafter, DOS). b, DOS binding site to Sestrin 2 predicted by AutoDock Vina and AlphaFold algorithms. c, Co-Immunoprecipitation of biotinylated DOS46L (DOS; used at 10μM, throughout) followed by recombinant tagged protein detection, as indicated. d, Recombinant sMAC activity investigated by liquid scintillation. Cpm, counts per minutes. e, Half maximal inhibitory concentration (IC50) of DOS against recombinant 385 sestrin-AMPK complexes by in vitro kinase assays with SAMS peptides and ADP-glo kinase assessment (left); equilibrium dissociation constant (Kd) of sestrin bound biotinylated DOS assessed by ELISA with streptavidin antibodies (right). f, Quantification of selective DOS penetration in sMAC negative and sMAC positive primary human CD4+ T cells by flow cytometry, 30 minutes after DOS treatment (n = 7 donors). g, Individual channels of representative confocal imaging (n = 3 donors) of DOS (green) uptake in human CD4+ T cells. Scale bar, 5 μm. h, Co-Immunoprecipitation (IP, hereafter) of biotinylated DOS from human CD4+ T cell extracts after overnight treatment. Presence or absence of sestrin, p38, JNK, ERK (sMAC) or MIOS and WDR59 (GATOR) was confirmed by western blot in the IP. Whole cell lysates (WCL) controls are shown (same extracts; run in parallel). i, ERK, JNK and p38 MAPK kinase activity (autophosphorylation) among Tsen sestrin 2 immunoprecipitates (sMACs) in the presence or in the absence of DOS for 30 minutes (n = 5 donors). Kinase activity was detected by ELISA assays. j, Human Tsen were pre-treated with the proteasomal inhibitor MG132 (1μM) for 30 minutes followed by addition of DOS for 2 hours. Controls were not treated with DOS. Total lysates and sestrin 2 immunoprecipitates were then analysed by immunoblotting of ubiquitin and sestrin 2. H3, whole cell lysate control. k, Pooled ubiquitination data (n = 5 donors) for experiments as in (i). Data are shown as fold change to the untreated control (NO DOS, in the presence of MG132), set as 1. l, Time dependent assessment of sestrin expression (1, 2 and 3) by DOS in human Tsen (top) and AMPK phosphorylation in the same cells assessed by western blotting (bottom). Representative of 6 donors. In e, f, j, k, one-way Anova with Bonferroni postcorrection for multiple comparisons was performed. **P\u0026lt;0,01; ***P\u0026lt;0,001; ****P\u0026lt;0,0001. Error bars indicate SEM.\u003c/p\u003e","description":"","filename":"NCBFig.1.png","url":"https://assets-eu.researchsquare.com/files/rs-5180379/v1/261b21c6c8dc1b085aa112d2.png"},{"id":66297519,"identity":"ba076fee-0fcb-4edb-8deb-ee10c21c136a","added_by":"auto","created_at":"2024-10-10 04:04:14","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1005587,"visible":true,"origin":"","legend":"\u003cp\u003eDOS-driven T cell reprograming a, Senescence-associated b-galactosidase expression in DOS-treated (DOS-juvenated) or untreated Tsen. Cells were purified and cultured for one week in the presence of anti-CD3 (0.5 μg/mL) and rhIL2 (5 ng/mL), then stained to detect β-galactosidase activity. Representative image on the inverted phasecontrast microscope (left) and relative quantification (right, n = 8 donors). b, Population doublings of human Tsen (transduced with irrelevant scramble) and sestrin 411 null CD4+ Tsen (transduced by triple lentiviral depletion of sestrins) and cultured as in (a ,left). Donor-matched CD27+ CD28+ CD4+ T cells (herafter, Terl) were cultured in parallel but activated with anti-CD3 and anti-CD28 (n = 5 donors). Cells were cultured over two weeks with restimulation every 7 days. DOS-driven population doublings (right) were calculated as delta between DOS treated and DOS untreated T cells, with or without depletion of sestrins. c, DOS-juvenation of human CD4+ T cells. Terminally differentiated effector memory CD45RA- CD28- CD27- CD4+ T cells (hereafter, senescent TEM) were purified and cultured over 20 days, as in (a) At day 1 (18 hours), 7, and 21 cell phenotypes were assessed by flow cytometry. Quantifications of rejuvenated stem like (CD28+ CD45RA+ CCR7+ CD95+ CD62L+ TCF1+) among human CD4+ T cells are shown (left; n = 5 donors). Decay of senescent TEM and CD28- CD27- CD45RA+ CD4+ T cells (hereafter, senescent TEMRA) undergoing rejuvenation is shown (right). d, Adoptive transfer of DOS-juvenated T cells, experimental design. Donor T cells were derived from twenty-month-old mice 15 days after Fluad vaccination with or without DOS treatment (0.1 mg/Kg throughout), labeled with Cell Trace Violet (CTV) dye or congenic CD45.1 tracking, then transferred into young naïve CD45.2 recipients (3 months). In parallel, young mice (3 months) were used as young donor control. Recipient animals were rested for 28 days, then analyzed for donor T cell persistence and maintenance of stem phenotype after transfer. e, Maintenance of donor DOS-juvenated CD45.1 CD4+ T cells, their aged-matched controls, and that of young donor T cells, 28 days after transfer (day 43) in recipient mouse lymph nodes. Representative flow cytometry plots and poled data (n = 5 mice per group) are shown. f, Assessment of stem like transferred T cells (among CD45.1 CD44- CD62L+ CD95+ CD4+ T cells) and terminally differentiated cells (TE, among CD45.1 CD44- CD62L- CD4+ T cells) following adoptive transfer as in (d) (n = 5 mice per group). g, IL7R gene expression and (h) CD95 and Sca-1 mean fluorescent intensity (MFI; throughout) in stem cells derived from CD4+ CD45.1+ transferred stem T cells among lymph nodes of recipient CD45.2 mice, 28 days after transfer (n = 5 mice per group). i, Representative plots of CD45.1 transferred cells in quiescent state before and after transfer assessed by cycle related intra-nuclear Ki67 staining. Representative of n = 5 mice per group. j, G1 (Ki67+) to G0 (Ki67-) transition in stem like CD45.1 CD4+ T cells 437 before and after adoptive transfer as indicated (n = 5 mice per group). k, Assessment of T cell longevity following DOS439 juvenation. Cells from lymph nodes were stained using the Annexin-PI Apoptosis detection Kit 28 days after transfer. Representative FACS plot (Left) and quantification of dead CD45.1+ CD4+ transferred T cells (Right). Data are from n = 4 mice per group. l, DOS-juvenated T cell maintenance, in vivo model. In a, b (right) two tailed paired T test was used. In b (left), c, e-h, j k one-way Anova with Bonferroni post-correction for multiple comparisons was used, *p\u0026lt;0,05, **P\u0026lt;0,01; ***P\u0026lt;0,001; ****P\u0026lt;0,0001. Error bars indicate SEM.\u003c/p\u003e","description":"","filename":"NCBFig.2.png","url":"https://assets-eu.researchsquare.com/files/rs-5180379/v1/cb4613feb2813092041eda0a.png"},{"id":66297424,"identity":"25a3d7b4-214c-4dcc-8343-46cfac7716e2","added_by":"auto","created_at":"2024-10-10 03:56:14","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":894839,"visible":true,"origin":"","legend":"\u003cp\u003eReprogramed T cells possess prophylactic functions even without vaccination a, Adoptive transfer of DOS-juvenated T cells, experimental design. CD45.1+ CD4+ donor T cells were derived from twenty-month-old mice injected with and without DOS (0.1 mg/Kg throughout), then transferred into old naïve CD45.2 recipients (20 months) 72 hours later. In parallel, young (3 month old) naïve CD45.1 mice were used as young CD4+ T cell donor control. Recipient animals were vaccinated, or not, with Fluad the day after, as indicated. Animals were subjected to a lethal H1N1 flu viral infection 8 weeks after transfer, and culled within 15 days of infection. Vaccinated young and old mice were also assessed, to determine background responses. b, Survival of mice, Fluad vaccinated, or not, and then challenged with lethal H1N1 infection as in (a). Data are from n = 5 mice per group. c, Fluad-specific IgG production in the same animals subjected to H1N1 infection. Data are from n = 5 mice per group. Transfer free vaccinated recipients are shown (top right; throughout). d, Neutralization properties of Flu-specific IgGs from recipient mice as indicated were assessed upon overnight H1N1 infection of canine kidney cells (MDCK) in vitro by ELISA with anti-influenza A NP antibody. Data are from n = 5 mice per group. e, Viral polymerase acidic protein (PA) assessment in lungs of the same mice upon H1N1 viral infection as in (a) by qPCR (n=5 mice per group). Data are expressed as fold change to the old, infected control (transferred CD4+ T cells, 462 no DOS), set as 1. f, Representative FACS plots and (g) pooled data of primary antiviral Th1 responses among T-bet+ IFNγ+ CD45.1 donor mouse CD4+ T cells as in (a). Data are from n = 5 mice per group. h, CD44- CD62L+ CD95+ CD4+ T cells (stem-like T cells) in animals subjected or not to CD4+ T cell transfer and then infected in the same experiments as in (a); Old naive recipient examples are shown (n = 5 mice per group). i, The fraction of sestrin expressing donor T cells among adoptively transferred CD45.1+ CD4+ T cells from age-matched (20 months) CD45.2+ recipients, six months after transfer as assessed by flow cytometry (n = 5 mice per group). In c-e, g-i, one-way Anova with Bonferroni post-correction for multiple comparisons was used. In c (Top right), e (Top right), a two tailed paired T test was used. In b Mantel-Cox test was used. *P\u0026lt;0,05; **P\u0026lt;0,01; ***P\u0026lt;0,001; ****P\u0026lt;0,0001. Error bars indicate SEM.\u003c/p\u003e","description":"","filename":"NCBFig.3.png","url":"https://assets-eu.researchsquare.com/files/rs-5180379/v1/680e3bcae46119ba42d8d9de.png"},{"id":66297429,"identity":"459bcf80-9349-4b42-84f9-1a2f301f433e","added_by":"auto","created_at":"2024-10-10 03:56:14","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":625845,"visible":true,"origin":"","legend":"\u003cp\u003eReprogramed T cells present de novo antigen-specific TCR rearrangements a, Purified human CD4+ T cells and autologous APCs (CD3-depleted PBMCs) were pre-treated or not with DOS or YF peptides (YF) for 4 hours, respectively, then co-cultured for additional 18 hours, in 1:2 ratio. The day after, post-synaptic CD4+ T cell genomic DNA was derived from the conjugates and processed for Next generation sequencing (NGS) of the TCRVB gene. The table shows inductions of T cell receptor Beta Variant (TRBV) rearrangements (V-DJ recombination) from a single donor upon YF stimulation of DOS-juvenated CD4+ T cells. For experimental details, please see Methods. One representative donor of 4 is shown. Further genomic clonotypes, Extended Data Fig.14. b, Antigen pocket rearrangements among TRBV7-1 variant in T cells derived from the same donor as in (a). Example of single YF-specific TCR clonotypes (among TRBV7-1) that are only seen in T cells exposed to YF-pulsed APCs, with or without DOS (bottom). Note that most antigen pocket rearranging requires presence of DOS, indicating active D-J recombination. c, Single cell presence of YF-specific TRBVs among the different T cell subsets of the 4 donors subjected to NGS. The expression levels of each TRBV were evaluated in individual cells by flow cytometry against the unstimulated background. Each dot is a single TRBV in the single cell flow cytometry assessment. No double TRBVs expressing T cells could be detected (top right). Results from 4 experiments are shown. d, Absence 488 of early YF-specific proliferation (18 hours) of the same stem T cells as shown by CTV tracking. YF-specific TRBV (7.1) clonotypes among the same stem-like T cell clones from 4 different donors, at the same time, is shown (intracellular flow IFN-γ detection; top right). Activation induced markers (AIM) assay of de novo TRBV7-1 YF-specific TSTEM clonotypes (n = 4 donors, bottom right). Antigen free synapses were subtracted. e, IFN-γ release by the indicated TRBV expressing T stem clones after 18-hour YF peptide stimulation and (f) antigen-specific proliferation of the same clones, 4 days after culture as assessed by BrdU incorporation. Cell culture conditions as in (a). Specificity cell population controls (TCM and TEM) are shown. In c, e One-way Anova with Bonferroni post-correction for multiple comparisons was used. In d, f statistical significance was determined by two tailed unpaired student T test. ***P\u0026lt;0,001; ****P\u0026lt;0,0001. Error bars indicate SEM.\u003c/p\u003e","description":"","filename":"NCBFig.4.png","url":"https://assets-eu.researchsquare.com/files/rs-5180379/v1/21dde3aafdea32557e2e4159.png"},{"id":66297439,"identity":"71bd5439-17e3-499f-8580-d504504ef76c","added_by":"auto","created_at":"2024-10-10 03:56:14","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":522149,"visible":true,"origin":"","legend":"\u003cp\u003eDOS restores Rag function via beta catenin during T cell reprograming a, Representative immunoblots (left) and quantification (right) of Rag protein expression from human CD4+ T cells treated or not with DOS for 4 hours (n = 3 donors). Data are expressed as fold change to the untreated control (no DOS), set as 1. b, Rag 2 proteins were immunoprecipitated from the chromatin of human CD4+ T cells treated as in (a) then followed by ELISA with anti-Rag 1 and 2 antibodies. Data are expressed as fold change to the untreated control (no DOS), set as 1. c and d, DOS driven Rag induction in the presence or in the absence of beta-catenin inhibition. Senescent TEM were pre-treated or not with the beta catenin inhibitor ICG-001 (10μM, 30’) followed by DOS driven reprograming for 4 hours. Rag expression was then investigated by quantitative real time PCR (c) and flow cytometry (d; n = 4 donors). Note absence of DOS-driven Rag generation in the presence of beta catenin blocking. e, YF-specific LCK activation (T394 phosphorylation) among senescent forming TEMRA cultured in the presence or in the absence of beta catenin (ICG-001) or Rag inhibition (Rag- by siRag1/2). Senescent TEM were pre-treated with beta-catenin or Rag inhibition then incubated with DOS for 4 hours, followed by conjugation with autologous YF pulsed APCs for 5 minutes. Results were 514 stratified by relative CD3 expression. Note that only the forming CD3low TEMRA undergo initial YF specific activation (n = 5 donors). f, Representative FACS plot validation (left) and quantification (right; n = 3 donors) of Rag knockdown in human CD4+ T cells. HEK293T cells were used as negative Rag expression control (bottom left). Note that Rag silencing reduces Rag expression to background fluorescence of HEK293 cells naturally lacking Rag proteins, validating known down efficiency. g, Rag deficiency abrogates DOS-driven TCR revisions. Human CD4+ T cells were transfected with siCtrl or siRAG1/siRAG2 for 3 days. The T cells were then treated with or without DOS for 4 hours, in the presence or in the absence of beta-catenin blocking (ICG-001) prior to culture with YF-pulsed or antigen-free APCs for 18 hours. The YF specific DOS-juvenated TCRs derived by background subtraction with no YF peptides are shown (n = 4 donors). h, Effect of Rag protein silencing on the YF-specific IFN-γ production among the indicated stem like T cell clones or on (i), the YF specific TCR rearrangement, assessed by TREC quantification (n = 4 donors). In a-d two tailed paired T test was used. In e-h One-way Anova with Bonferroni correction for multiple comparisons. In f, F test. *P\u0026lt;0,05; **P\u0026lt;0,01; ***P\u0026lt;0,001; ****P\u0026lt;0,0001. 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M.K. serves as a scientific advisor to Sentcell ltd. A.L. M.C and F.R. are supported by Sentcell ltd.","formattedTitle":"Rejuvenation driven reprograming in T lymphocytes","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":"","lastPublishedDoi":"10.21203/rs.3.rs-5180379/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5180379/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Rejuvenation restores cellular function lost with age, but it is not known to confer properties the cell lacked in its younger past. We found that Disruptors of sestrin-MAPK interactions (DOS) reprogramed senescent T cells into new stem like T clones with different T cell receptors (TCRs). DOS targeted sestrin-MAPK complexes (sMAC) to ubiquitin-dependent proteasomal degradation, resulting in long-term sestrin transcriptional inhibition, increased T cell fitness, and generation of long-lived stem like memory clones. Strikingly, DOS reactivated juvenile related Rag re-expression, leading to antigen-specific TCR rearrangements in formerly senescent cells. As such, rejuvenated T cells, with stem features and new TCRs, combated multiple lethal challenges never previously encountered in their lifetime, with no need for vaccination. T cell rejuvenation confers new properties whilst preserving cellular state, which unravels an unknown rejuvenating principle in biological systems.","manuscriptTitle":"Rejuvenation driven reprograming in T lymphocytes","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-10-10 03:56:09","doi":"10.21203/rs.3.rs-5180379/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":"dc75be64-4a26-475d-8f0c-45778138e3e1","owner":[],"postedDate":"October 10th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":38647923,"name":"Biological sciences/Immunology/Signal transduction"},{"id":38647924,"name":"Biological sciences/Immunology/Lymphocytes/T cells"}],"tags":[],"updatedAt":"2025-03-18T20:30:42+00:00","versionOfRecord":[],"versionCreatedAt":"2024-10-10 03:56:09","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5180379","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5180379","identity":"rs-5180379","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}