{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,24]],"date-time":"2026-07-24T12:27:59Z","timestamp":1784896079456,"version":"3.55.0"},"reference-count":40,"publisher":"Oxford University Press (OUP)","issue":"4","license":[{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"DOI":"10.13039\/501100001807","name":"S\u00e3o Paulo Research Foundation","doi-asserted-by":"publisher","award":["17\/10780\u20134"],"award-info":[{"award-number":["17\/10780\u20134"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"National Council for Scientific and Technological Development","doi-asserted-by":"publisher","award":["311304\/2021"],"award-info":[{"award-number":["311304\/2021"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"National Council for Scientific and Technological Development","doi-asserted-by":"publisher","award":["302060\/2019\u20137"],"award-info":[{"award-number":["302060\/2019\u20137"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"name":"CAPES-STINT","award":["88887.642780\/2021\u201300"],"award-info":[{"award-number":["88887.642780\/2021\u201300"]}]},{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","award":["001"],"award-info":[{"award-number":["001"]}],"id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,2,23]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Autoimmune Polyendocrine Syndrome Type 1 (APS-1) is a rare monogenic disorder caused by mutations in the autoimmune regulator (AIRE) gene. Although AIRE is essential for central immune tolerance, how distinct APS-1\u2013associated mutations differentially affect medullary thymic epithelial cell (mTEC) biology remains incompletely understood. Here, we investigated the molecular and functional consequences of three Aire\/AIRE variants using complementary murine mTEC models. To define transcriptional effects, we performed single-cell RNA sequencing (scRNA-seq) on mTECs carrying a heterozygous genomic Aire c.735delG mutation generated by CRISPR-Cas9. This analysis revealed reduced transcriptional heterogeneity, decreased expression of tissue-restricted antigens (TRAs) mRNAs (including Col4a3, Col7a1, and Neto2), and downregulation of key mTEC lineage markers (Epcam, Cldn4, Krt14). Mutant cells also displayed altered expression of mRNAs involved in chemokine-mediated migration (Ccl25, Cxcl16), extracellular matrix and cell adhesion (Fn1, Lama5, Col4a1, Nectin1, Cdh1), and actin cytoskeleton organization (Gsn, Rac1, Wasl, Actn1), indicating broad disruption of pathways governing mTEC identity and cell\u2013cell interactions. Guided by these findings, we assessed mutation-specific functional outcomes using a CRISPR-derived Aire functional knockout and lentiviral expression of the human AIRE missense variants p.G229W and p.C313Y in wild-type mTECs. Functional assays revealed mutation-dependent alterations in mTEC morphology, thymocyte migration, and adhesion, with the p.C313Y variant exerting the strongest effects. Together, these data demonstrate that heterozygous and missense AIRE mutations exert distinct yet convergent effects on mTEC transcriptional programs and cellular behavior, providing mechanistic insight into AIRE-dependent immune tolerance failure in APS-1.<\/jats:p>","DOI":"10.1093\/hmg\/ddag004","type":"journal-article","created":{"date-parts":[[2026,1,28]],"date-time":"2026-01-28T12:41:43Z","timestamp":1769604103000},"source":"Crossref","is-referenced-by-count":1,"title":["Distinct mutations in the autoimmune regulator gene differentially affect transcriptional and functional properties of medullary thymic epithelial cells"],"prefix":"10.1093","volume":"35","author":[{"given":"Pedro Paranhos","family":"Tanaka","sequence":"first","affiliation":[{"name":"Molecular Immunogenetics Group, Department of Genetics, Ribeir\u00e3o Preto Medical School, University of S\u00e3o Paulo , 3900, Bandeirantes Avenue, 14040-900, Ribeir\u00e3o Preto, SP ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vinicius","family":"Cotta-Almeida","sequence":"additional","affiliation":[{"name":"Laboratory on Thymus Research, Oswaldo Cruz Foundation, Fiocruz , 4365, Brasil Avenue, Rio de Janeiro, RJ, 21040-360 ,","place":["Brazil"]},{"name":"National Institute of Science and Technology on Neuroimmunomodulation, INCT-NIM, Fiocruz , 4365, Brasil Avenue, Rio de Janeiro, RJ, 21040-360 ,","place":["Brazil"]},{"name":"Rio de Janeiro Research Network on Neuroinflammation, RENEURIN, Fiocruz , 4365, Brasil Avenue, Rio de Janeiro, RJ, 21040-360 ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eduardo Ant\u00f4nio","family":"Donadi","sequence":"additional","affiliation":[{"name":"Department of Clinical Medicine, Division of Clinical Immunology, Ribeir\u00e3o Preto Medical School, University of S\u00e3o Paulo , 3900, Bandeirantes Avenue, 14040-900, Ribeir\u00e3o Preto, SP ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa","family":"Westerberg","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet , Nobels v\u00e4g 5, 17165 Stockholm ,","place":["Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4408-140X","authenticated-orcid":false,"given":"Geraldo Aleixo","family":"Passos","sequence":"additional","affiliation":[{"name":"Molecular Immunogenetics Group, Department of Genetics, Ribeir\u00e3o Preto Medical School, University of S\u00e3o Paulo , 3900, Bandeirantes Avenue, 14040-900, Ribeir\u00e3o Preto, SP ,","place":["Brazil"]},{"name":"Laboratory of Genetics and Molecular Biology, Department of Basic and Oral Biology, Ribeir\u00e3o Preto School of Dentistry, University of S\u00e3o Paulo , Via do Caf\u00e9 s\/n, Ribeir\u00e3o Preto, SP, 14040-904 ,","place":["Brazil"]},{"name":"Center for Cell-Based Therapy in Dentistry, Ribeir\u00e3o Preto School of Dentistry, University of S\u00e3o Paulo , Via do Caf\u00e9 s\/n, Ribeir\u00e3o Preto, SP, 14040-904 ,","place":["Brazil"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,2,13]]},"reference":[{"key":"2026051101414885300_ref1","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1146\/annurev-immunol-051116-052320","article-title":"Thymic epithelial cells","volume":"35","author":"Abramson","year":"2017","journal-title":"Annu Rev Immunol"},{"key":"2026051101414885300_ref2","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1146\/annurev-immunol-101320-022432","article-title":"Instructive cues of thymic T cell selection","volume":"40","author":"Irla","year":"2022","journal-title":"Annu Rev Immunol"},{"key":"2026051101414885300_ref3","doi-asserted-by":"publisher","first-page":"1376655","DOI":"10.3389\/fimmu.2024.1376655","article-title":"The single-cell transcriptome of mTECs and CD4+ thymocytes under adhesion revealed heterogeneity of mTECs and a network controlled by Aire and lncRNAs","volume":"15","author":"Monteiro","year":"2024","journal-title":"Front Immunol"},{"key":"2026051101414885300_ref4","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1007\/978-3-031-77921-3_3","article-title":"The ins and outs of thymic epithelial cell differentiation and function","volume":"1471","author":"Matsumoto","year":"2025","journal-title":"Adv Exp Med Biol"},{"key":"2026051101414885300_ref5","doi-asserted-by":"publisher","first-page":"4296","DOI":"10.1038\/s41467-022-31750-1","article-title":"Transcriptomic diversity in human medullary thymic epithelial cells","volume":"13","author":"Carter","year":"2022","journal-title":"Nat Commun"},{"key":"2026051101414885300_ref6","doi-asserted-by":"publisher","first-page":"948259","DOI":"10.3389\/fimmu.2022.948259","article-title":"Aire and Fezf2, two regulators in medullary thymic epithelial cells, control autoimmune diseases by regulating TSAs: partner or complementer?","volume":"13","author":"Qi","year":"2022","journal-title":"Front Immunol"},{"key":"2026051101414885300_ref7","doi-asserted-by":"publisher","first-page":"10","DOI":"10.1111\/imm.12831","article-title":"Update on Aire and thymic negative selection","volume":"153","author":"Passos","year":"2018","journal-title":"Immunology"},{"key":"2026051101414885300_ref8","doi-asserted-by":"publisher","first-page":"2827","DOI":"10.1084\/jem.20080046","article-title":"Aire controls the differentiation program of thymic epithelial cells in the medulla for the establishment of self-tolerance","volume":"205","author":"Yano","year":"2008","journal-title":"J Exp Med"},{"key":"2026051101414885300_ref9","doi-asserted-by":"publisher","first-page":"975","DOI":"10.1016\/j.cell.2015.10.013","article-title":"Fezf2 orchestrates a thymic program of self-antigen expression for immune 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