{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,12]],"date-time":"2026-03-12T01:05:35Z","timestamp":1773277535112,"version":"3.50.1"},"reference-count":47,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,12,20]],"date-time":"2021-12-20T00:00:00Z","timestamp":1639958400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100002608","name":"American Thyroid Association","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100002608","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:p>Autoimmune polyglandular syndrome type 3 variant (APS3v) refers to an autoimmune condition in which both type 1 diabetes (T1D) and autoimmune thyroiditis (AITD) develop in the same individual. HLA-DR3 confers the strongest susceptibility to APS3v. Previously we reported a unique amino acid signature pocket that predisposes to APS3v. We found that this pocket is flexible and can trigger APS3v by presenting both thyroid (Tg.1571, TPO.758) and islet (GAD.492) peptides to induce autoimmune response. We hypothesized that blocking the specific APS3v-HLA-DR3 pocket from presenting thyroid\/islet antigens can block the autoimmune response in APS3v. To test this hypothesis we performed a virtual screen of small molecules blocking APS3v-HLA-DR3, and identified 11 small molecules hits that were predicted to block APS3v-HLA-DR3. Using the baculovirus-produced recombinant APS3v-HLA-DR3 protein we tested the 11 small molecules in an <jats:italic>in vitro<\/jats:italic> binding assay. We validated 4 small molecule hits, S9, S5, S53 and S15, that could block the APS3v-HLA-DR3 pocket <jats:italic>in vitro<\/jats:italic>. We then developed a novel humanized APS3v mouse model induced by co-immunizing a peptide mix of Tg.1571, TPO.758 and GAD.492. The immunized mice developed strong T-cell and antibody responses to the thyroid\/islet peptides, as well as mouse thyroglobulin. In addition, the mice showed significantly lower free T4 levels compared to controls. Using the APS3v mouse model, we showed that one of the 4 small molecules, Cepharanthine (S53), blocked T-cell activation by thyroid\/islet peptides <jats:italic>ex vivo<\/jats:italic> and <jats:italic>in vivo<\/jats:italic>. These findings suggested Cepharanthine may have a therapeutic potential in APS3v patients carrying the specific APS3v-HLA-DR3 pocket.<\/jats:p>","DOI":"10.3389\/fimmu.2021.796552","type":"journal-article","created":{"date-parts":[[2021,12,20]],"date-time":"2021-12-20T06:58:16Z","timestamp":1639983496000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["Cepharanthine Blocks Presentation of Thyroid and Islet Peptides in a Novel Humanized Autoimmune Diabetes and Thyroiditis Mouse Model"],"prefix":"10.3389","volume":"12","author":[{"given":"Cheuk Wun","family":"Li","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Roman","family":"Osman","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesca","family":"Menconi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Larissa C.","family":"Faustino","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kookjoo","family":"Kim","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Oliver B.","family":"Clarke","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hanxi","family":"Hou","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yaron","family":"Tomer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2021,12,20]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1006\/clin.1997.4412","article-title":"Epidemiology and Estimated Population Burden of Selected Autoimmune Diseases in the United States","volume":"84","author":"Jacobson","year":"1997","journal-title":"Clin Immunol Immunopathol"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.2337\/diacare.23.10.1516","article-title":"Incidence of Childhood Type 1 Diabetes Worldwide. Diabetes Mondiale (DiaMond) Project Group","volume":"23","author":"Karvonen","year":"2000","journal-title":"Diabetes Care"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1089\/105072502320908312","article-title":"Disease Associations With Autoimmune Thyroid Disease","volume":"12","author":"Jenkins","year":"2002","journal-title":"Thyroid"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMra030158","article-title":"Autoimmune Polyendocrine Syndromes","volume":"350","author":"Eisenbarth","year":"2004","journal-title":"N Engl J Med"},{"key":"B5","doi-asserted-by":"publisher","first-page":"697","DOI":"10.1210\/er.2008-0015","article-title":"Joint Genetic Susceptibility to Type 1 Diabetes and Autoimmune Thyroiditis: From Epidemiology to Mechanisms","volume":"29","author":"Huber","year":"2008","journal-title":"Endocr Rev"},{"key":"B6","first-page":"9","article-title":"Update on Autoimmune Polyendocrine Syndromes (APS)","volume":"74","author":"Betterle","year":"2003","journal-title":"Acta BioMed"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1210\/jc.2004-2236","article-title":"Genetic Analysis of Families With Autoimmune Diabetes and Thyroiditis: Evidence for Common and Unique Genes","volume":"90","author":"Golden","year":"2005","journal-title":"J Clin Endocrinol Metab"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1210\/jc.2008-2193","article-title":"Autoimmune Thyroiditis and Diabetes: Dissecting the Joint Genetic Susceptibility in a Large Cohort of Multiplex Families","volume":"94","author":"Villano","year":"2009","journal-title":"J Clin Endocrinol Metab"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1089\/thy.2010.1639","article-title":"Genetics of the Autoimmune Polyglandular Syndrome Type 3 Variant","volume":"20","author":"Dittmar","year":"2010","journal-title":"Thyroid"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1009511107","article-title":"Shared Molecular Amino Acid Signature in the HLA-DR Peptide Binding Pocket Predisposes to Both Autoimmune Diabetes and Thyroiditis","volume":"107","author":"Menconi","year":"2010","journal-title":"Proc Natl Acad Sci USA"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaut.2015.03.006","article-title":"Genome Wide Identification of New Genes and Pathways in Patients With Both Autoimmune Thyroiditis and Type 1 Diabetes","volume":"60","author":"Tomer","year":"2015","journal-title":"J Autoimmun"},{"key":"B12","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.jaut.2016.09.007","article-title":"Flexible Peptide Recognition by HLA-DR Triggers Specific Autoimmune T-Cell Responses in Autoimmune Thyroiditis and Diabetes","volume":"76","author":"Li","year":"2017","journal-title":"J Autoimmun"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M109.041574","article-title":"Employing a Recombinant HLA-DR3 Expression System to Dissect Major Histocompatibility Complex II-Thyroglobulin Peptide Dynamism: A Genetic, Biochemical, and Reverse Immunological Perspective","volume":"284","author":"Jacobson","year":"2009","journal-title":"J Biol Chem"},{"key":"B14","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1084\/jem.178.1.27","article-title":"Specificity and Promiscuity Among Naturally Processed Peptides Bound to HLA-DR Alleles","volume":"178","author":"Chicz","year":"1993","journal-title":"J Exp Med"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M115.694687","article-title":"Identifying a Small Molecule Blocking Antigen Presentation in Autoimmune Thyroiditis","volume":"291","author":"Li","year":"2016","journal-title":"J Biol Chem"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1021\/jm051256o","article-title":"Extra Precision Glide: Docking and Scoring Incorporating a Model of Hydrophobic Enclosure for Protein-Ligand Complexes","volume":"49","author":"Friesner","year":"2006","journal-title":"J Med Chem"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1007\/s002510050277","article-title":"Role of Mouse and Human Class II Transgenes in Susceptibility to and Protection Against Mouse Autoimmune Thyroiditis","volume":"46","author":"Kong","year":"1997","journal-title":"Immunogenetics"},{"key":"B18","doi-asserted-by":"publisher","first-page":"102402","DOI":"10.1016\/j.jaut.2020.102402","article-title":"Cepharanthine Blocks TSH Receptor Peptide Presentation by HLA-DR3: Therapeutic Implications to Graves' Disease","volume":"108","author":"Li","year":"2020","journal-title":"J Autoimmun"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1016\/j.lfs.2006.12.001","article-title":"The Effects of Biscoclaurine Alkaloid Cepharanthine on Mammalian Cells: Implications for Cancer, Shock, and Inflammatory Diseases","volume":"80","author":"Furusawa","year":"2007","journal-title":"Life Sci"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1016\/S1734-1140(11)70500-X","article-title":"Therapeutic Potential of the Biscoclaurine Alkaloid, Cepharanthine, for a Range of Clinical Conditions","volume":"63","author":"Rogosnitzky","year":"2011","journal-title":"Pharmacol Rep"},{"key":"B21","doi-asserted-by":"publisher","first-page":"152956","DOI":"10.1016\/j.phymed.2019.152956","article-title":"Cepharanthine: An Update of its Mode of Action, Pharmacological Properties and Medical Applications","volume":"62","author":"Bailly","year":"2019","journal-title":"Phytomedicine"},{"key":"B22","first-page":"47","article-title":"Effect of Cepharanthine on Alopecia Areata","volume":"64","author":"Niwa","year":"1969","journal-title":"Hifuka Kiyo"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1111\/j.1346-8138.2002.tb00199.x","article-title":"Seventeen Cases of Alopecia Areata: Combination of SADBE Topical Immunotherapy With Other Therapies","volume":"29","author":"Morita","year":"2002","journal-title":"J Dermatol"},{"key":"B24","article-title":"Effects of Cepharanthin on Leukopenia and Thrombocytopenia Caused by CDDP-ACR-CPA Therapy of Ovarian Cancer","volume":"15","author":"Ushiki","year":"1988","journal-title":"Gan To Kagaku Ryoho"},{"key":"B25","article-title":"Clinical Effects of Cepharanthin (Ceph.) on Leukopenia by Chemotherapy in Lung Cancer Patients","volume":"24","author":"Saito","year":"1989","journal-title":"Nihon Gan Chiryo Gakkai Shi"},{"key":"B26","article-title":"Effect of Cepharanthin on Radiotherapy Induced Leukopenia","volume":"35","author":"Ohta","year":"1990","journal-title":"Rinsho Hoshasen"},{"key":"B27","article-title":"Effects of Cepharanthin on Leukopenia and Thrombocytopenia Induced by Chemotherapy in Lung Cancer Patients","volume":"19","author":"Suzuki","year":"1992","journal-title":"Gan To Kagaku Ryoho"},{"key":"B28","article-title":"Therapeutic Experience of Venomous Snakebites by the Japanese Viper (Agkistrodon Halys Blomhoffii) With Low Dose of Antivenin: Report of 43 Consecutive Cases","volume":"66","author":"Kimoto","year":"1997","journal-title":"Nihon Geka Hokan"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1016\/j.intimp.2012.04.015","article-title":"Low Dose Cepharanthine Ameliorates Immune Thrombocytopenic Purpura Associated With Multiple Myeloma","volume":"13","author":"Tabata","year":"2012","journal-title":"Int Immunopharmacol"},{"key":"B30","article-title":"Antitumor Effect of Cepharanthin in the Double Grafted Tumor System","volume":"17","author":"Ebina","year":"1990","journal-title":"Gan To Kagaku Ryoho"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1016\/0006-2952(91)90012-T","article-title":"Inhibition of 12-O-Tetradecanoyl Phorbol-13-Acetate Promoted Tumorigenesis by Cepharanthine, a Biscoclaurine Alkaloid, in Relation to the Inhibitory Effect on Protein Kinase C","volume":"41","author":"Edashige","year":"1991","journal-title":"Biochem Pharmacol"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.3892\/ijo_00000417","article-title":"Cepharanthine Inhibits Angiogenesis and Tumorigenicity of Human Oral Squamous Cell Carcinoma Cells by Suppressing Expression of Vascular Endothelial Growth Factor and Interleukin-8","volume":"35","author":"Harada","year":"2009","journal-title":"Int J Oncol"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1254\/jjp.54.464","article-title":"Reversal of Resistance to Doxorubicin With Cepharanthine in Murine P388 Leukemia Cells","volume":"54","author":"Fujimura","year":"1990","journal-title":"Jpn J Pharmacol"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1111\/j.1349-7006.2003.tb01482.x","article-title":"Reversal of the Resistance to STI571 in Human Chronic Myelogenous Leukemia K562 Cells","volume":"94","author":"Mukai","year":"2003","journal-title":"Cancer Sci"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1254\/fpj.88.71","article-title":"Effects of Cepharanthine on Experimental Nasal Allergy","volume":"88","author":"Kohno","year":"1986","journal-title":"Nihon Yakurigaku Zasshi"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1254\/fpj.90.205","article-title":"[Mode of the Anti-Allergic Action of Cepharanthine on an Experimental Model of Allergic Rhinitis]","volume":"90","author":"Kohno","year":"1987","journal-title":"Nihon Yakurigaku Zasshi"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1111\/ped.13151","article-title":"High-Dose Cepharanthin for Pediatric Chronic Immune Thrombocytopenia in Japan","volume":"59","author":"Yamazaki","year":"2017","journal-title":"Pediatr Int"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0806584105","article-title":"Molecular Amino Acid Signatures in the MHC Class II Peptide-Binding Pocket Predispose to Autoimmune Thyroiditis in Humans and in Mice","volume":"105","author":"Menconi","year":"2008","journal-title":"Proc Natl Acad Sci USA"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.2174\/138920207783591690","article-title":"The HLA Region and Autoimmune Disease: Associations and Mechanisms of Action","volume":"8","author":"Gough","year":"2007","journal-title":"Curr Genomics"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1210\/jc.2013-2852","article-title":"HLA Class II Haplotypes Differentiate Between the Adult Autoimmune Polyglandular Syndrome Types II and III","volume":"99","author":"Flesch","year":"2014","journal-title":"J Clin Endocrinol Metab"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.2337\/db11-0364","article-title":"Genetic Analysis of Adult-Onset Autoimmune Diabetes","volume":"60","author":"Howson","year":"2011","journal-title":"Diabetes"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1007\/s00125-015-3514-y","article-title":"The 6 Year Incidence of Diabetes-Associated Autoantibodies in Genetically at-Risk Children: The TEDDY Study","volume":"58","author":"Krischer","year":"2015","journal-title":"Diabetologia"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.2337\/dc17-0238","article-title":"Genetic and Environmental Interactions Modify the Risk of Diabetes-Related Autoimmunity by 6 Years of Age: The TEDDY Study","volume":"40","author":"Krischer","year":"2017","journal-title":"Diabetes Care"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1007\/s00125-017-4308-1","article-title":"Early Prediction of Autoimmune (Type 1) Diabetes","volume":"60","author":"Regnell","year":"2017","journal-title":"Diabetologia"},{"key":"B45","doi-asserted-by":"publisher","first-page":"591","DOI":"10.1007\/s00125-020-05344-9","article-title":"First-Appearing Islet Autoantibodies for Type 1 Diabetes in Young Children: Maternal Life Events During Pregnancy and the Child's Genetic Risk","volume":"64","author":"Johnson","year":"2021","journal-title":"Diabetologia"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1172\/JCI97739","article-title":"Methyldopa Blocks MHC Class II Binding to Disease-Specific Antigens in Autoimmune Diabetes","volume":"128","author":"Ostrov","year":"2018","journal-title":"J Clin Invest"},{"key":"B47","doi-asserted-by":"publisher","first-page":"102543","DOI":"10.1016\/j.jaut.2020.102543","article-title":"Retro-Inverso D-Peptides as a Novel Targeted Immunotherapy for Type 1 Diabetes","volume":"115","author":"Lombardi","year":"2020","journal-title":"J Autoimmun"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2021.796552\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,12,20]],"date-time":"2021-12-20T06:58:22Z","timestamp":1639983502000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2021.796552\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,20]]},"references-count":47,"alternative-id":["10.3389\/fimmu.2021.796552"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2021.796552","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,12,20]]},"article-number":"796552"}}