{"paper_id":"4572ece3-9145-4602-b3a1-4b4e2cbf7fee","body_text":"The cooperation between RNA binding proteins ZFP36L1 and ZFP36L2 controls thymic epithelial cell differentiation to promote thymus function. | 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 The cooperation between RNA binding proteins ZFP36L1 and ZFP36L2 controls thymic epithelial cell differentiation to promote thymus function. Nuno Alves, Pedro Ferreirinha, Pedro Rodrigues, Francisco Sobral, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6941685/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 The thymus generates immunocompetent T cells, but its function declines with age. Thymic activity depends on the proper differentiation of thymic epithelial cells (TECs), which establish unique niches for T-cell development. However, the molecular mechanisms governing the differentiation of functionally diverse cortical (c) and medullary (m) TEC subsets from common progenitors remain poorly understood. Employing dual conditional knockout (dcKO) and lineage-tracing mouse models, we demonstrate that the RNA-binding proteins ZFP36L1 and ZFP36L2 cooperatively maintain a functional TEC microenvironment. Deletion of Zfp36l1 and Zfp36l2 in TEC-specific dcKO mouse models resulted in similarly pronounced and early-onset thymic hypoplasia, with a significant reduction in both TEC and thymocyte cellularity. Mechanistically, single-cell transcriptomics showed significant alterations in the composition and transcriptional landscape of cTECs, mTECs, and thymic mimetic cells of dcKO thymus. The erosion of the TEC compartment, together with the accumulation of age- associated TECs suggested impaired differentiation and maintenance of mature TEC lineages from their progenitors. Fate-mapping analysis revealed that dual deficiency in Zfp36l1 and Zfp36l2 disrupted the canonical developmental trajectory from b5t-expressing TEC progenitors into mature cTEC and mTEC lineages. Our findings uncover a cooperative role for ZFP36L1 and ZFP36L2 in orchestrating TEC lineage differentiation and determining thymic function under physiological conditions. Biological sciences/Immunology/Lymphoid tissues/Thymus Biological sciences/Immunology/Adaptive immunity/Cellular immunity/Lymphocyte differentiation Full Text Additional Declarations All experiments requiring the use of mouse models (Mus muscles) were performed in accordance with European guidelines according with the directive 2010/63/EU and approved by Institutional Review Board and the Portuguese national authority for animal experimentation (DGAV) - Directorate General for Food and Veterinary Ministry of Agriculture. Project: Molecular Control of self-renewal and lineage specification in thymic epithelial cell progenitors in vivo; license number 008807/2021-06-22 There is NO Competing Interest. Supplementary Files SupFigures.pdf Supplemental Figures 1-6 Supp.Table1.xlsx Supplemental table 1 Supp.Table2.xlsx Supplemental table 2 Supp.Table3.xlsx Supplemental table 3 Supp.Table4.GOBPupregulated.xlsx Supplemental table 4 GO_BP Upregulated 2.Supp.Table4.GOBPDownregulated.xlsx Supplemental table 4 GO_BP Downpregulated 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. 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-6941685\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":true,\"archivedVersions\":[],\"articleType\":\"Article\",\"associatedPublications\":[],\"authors\":[{\"id\":475976381,\"identity\":\"83c7bba0-830f-4c26-b606-b87ac9908005\",\"order_by\":0,\"name\":\"Nuno 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Thymic activity depends on the proper differentiation of thymic epithelial cells (TECs), which establish unique niches for T-cell development. However, the molecular mechanisms governing the differentiation of functionally diverse cortical (c) and medullary (m) TEC subsets from common progenitors remain poorly understood. Employing dual conditional knockout (dcKO) and lineage-tracing mouse models, we demonstrate that the RNA-binding proteins ZFP36L1 and ZFP36L2 cooperatively maintain a functional TEC microenvironment. Deletion of Zfp36l1 and Zfp36l2 in TEC-specific dcKO mouse models resulted in similarly pronounced and early-onset thymic hypoplasia, with a significant reduction in both TEC and thymocyte cellularity. Mechanistically, single-cell transcriptomics showed significant alterations in the composition and transcriptional landscape of cTECs, mTECs, and thymic mimetic cells of dcKO thymus. The erosion of the TEC compartment, together with the accumulation of age- associated TECs suggested impaired differentiation and maintenance of mature TEC lineages from their progenitors. Fate-mapping analysis revealed that dual deficiency in Zfp36l1 and Zfp36l2 disrupted the canonical developmental trajectory from b5t-expressing TEC progenitors into mature cTEC and mTEC lineages. 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