{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,24]],"date-time":"2026-03-24T22:45:43Z","timestamp":1774392343831,"version":"3.50.1"},"reference-count":35,"publisher":"Oxford University Press (OUP)","issue":"8","license":[{"start":{"date-parts":[[2016,5,5]],"date-time":"2016-05-05T00:00:00Z","timestamp":1462406400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"name":"Italian Ministry of Health","award":["GR-2009-1555694"],"award-info":[{"award-number":["GR-2009-1555694"]}]},{"name":"Italian Ministry of Health","award":["H31J11000260001"],"award-info":[{"award-number":["H31J11000260001"]}]},{"name":"Italian Ministry of Health","award":["GR-2010-2316138"],"award-info":[{"award-number":["GR-2010-2316138"]}]},{"name":"Italian Ministry of Health","award":["H41J12000100001"],"award-info":[{"award-number":["H41J12000100001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2016,8,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:sec>\n                  <jats:title\/>\n                  <jats:p>Ectrodactyly-ectodermal dysplasia-clefting (EEC) syndrome is a rare autosomal dominant disease caused by mutations in the p63 gene. To date, approximately 40 different p63 mutations have been identified, all heterozygous. No definitive treatments are available to counteract and resolve the progressive corneal degeneration due to a premature aging of limbal epithelial stem cells. Here, we describe a unique case of a young female patient, aged 18 years, with EEC and corneal dysfunction, who was, surprisingly, homozygous for a novel and de novo R311K missense mutation in the p63 gene. A detailed analysis of the degree of somatic mosaicism in leukocytes from peripheral blood and oral mucosal epithelial stem cells (OMESCs) from biopsies of buccal mucosa showed that approximately 80% were homozygous mutant cells and 20% were heterozygous. Cytogenetic and molecular analyses excluded genomic alterations, thus suggesting a de novo mutation followed by an allelic gene conversion of the wild-type allele by de novo mutant allele as a possible mechanism to explain the homozygous condition. R311K-p63 OMESCs were expanded in vitro and heterozygous holoclones selected following clonal analysis. These R311K-p63 OMESCs were able to generate well-organized and stratified epithelia in vitro, resembling the features of healthy tissues. This study supports the rationale for the development of cultured autologous oral mucosal epithelial stem cell sheets obtained by selected heterozygous R311K-p63 stem cells, as an effective and personalized therapy for reconstructing the ocular surface of this unique case of EEC syndrome, thus bypassing gene therapy approaches.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Significance<\/jats:title>\n                  <jats:p>This case demonstrates that in a somatic mosaicism context, a novel homozygous mutation in the p63 gene can arise as a consequence of an allelic gene conversion event, subsequent to a de novo mutation. The heterozygous mutant R311K-p63 stem cells can be isolated by means of clonal analysis and given their good regenerative capacity, they may be used to successfully correct the corneal defects present in this unique case of ectrodactyly-ectodermal dysplasia-clefting syndrome.<\/jats:p>\n               <\/jats:sec>","DOI":"10.5966\/sctm.2015-0358","type":"journal-article","created":{"date-parts":[[2016,5,6]],"date-time":"2016-05-06T04:19:31Z","timestamp":1462508371000},"page":"1098-1105","source":"Crossref","is-referenced-by-count":14,"title":["Personalized Stem Cell Therapy to Correct Corneal Defects Due to a Unique Homozygous-Heterozygous Mosaicism of Ectrodactyly-Ectodermal Dysplasia-Clefting Syndrome"],"prefix":"10.1093","volume":"5","author":[{"given":"Vanessa","family":"Barbaro","sequence":"first","affiliation":[{"name":"Fondazione Banca degli Occhi del Veneto, Venice, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annamaria Assunta","family":"Nasti","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padua, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paolo","family":"Raffa","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padua, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angelo","family":"Migliorati","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padua, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Patrizia","family":"Nespeca","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padua, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stefano","family":"Ferrari","sequence":"additional","affiliation":[{"name":"Fondazione Banca degli Occhi del Veneto, Venice, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elisa","family":"Palumbo","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Padua, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marina","family":"Bertolin","sequence":"additional","affiliation":[{"name":"Fondazione Banca degli Occhi del Veneto, Venice, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claudia","family":"Breda","sequence":"additional","affiliation":[{"name":"Fondazione Banca degli Occhi del Veneto, Venice, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesco","family":"Miceli","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, University of Naples Federico II, Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antonella","family":"Russo","sequence":"additional","affiliation":[{"name":"Department of Biology, University of Padua, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luciana","family":"Caenazzo","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padua, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Diego","family":"Ponzin","sequence":"additional","affiliation":[{"name":"Fondazione Banca degli Occhi del Veneto, Venice, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giorgio","family":"Pal\u00f9","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padua, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cristina","family":"Parolin","sequence":"additional","affiliation":[{"name":"Department of Molecular Medicine, University of Padua, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Enzo","family":"Di Iorio","sequence":"additional","affiliation":[{"name":"Fondazione Banca degli Occhi del Veneto, Venice, Italy"},{"name":"Department of Molecular Medicine, University of Padua, Padua, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2016,5,5]]},"reference":[{"key":"2021122705113476300_R1","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1086\/323123","article-title":"p63 Gene mutations in EEC syndrome, limb-mammary syndrome, and isolated split hand-split foot malformation suggest a genotype-phenotype correlation","volume":"69","author":"van Bokhoven","year":"2001","journal-title":"Am J Hum Genet"},{"key":"2021122705113476300_R2","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1111\/j.1365-2133.2009.09496.x","article-title":"Molecular basis of EEC (ectrodactyly, ectodermal dysplasia, clefting) syndrome: Five new mutations in the DNA-binding domain of the TP63 gene and genotype-phenotype correlation","volume":"162","author":"Clements","year":"2010","journal-title":"Br J Dermatol"},{"key":"2021122705113476300_R3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1086\/341450","article-title":"Splitting p63","volume":"71","author":"van Bokhoven","year":"2002","journal-title":"Am J Hum Genet"},{"key":"2021122705113476300_R4","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1016\/j.ophtha.2011.06.044","article-title":"Limbal stem cell deficiency and ocular phenotype in ectrodactyly-ectodermal dysplasia-clefting syndrome caused by p63 mutations","volume":"119","author":"Di Iorio","year":"2012","journal-title":"Ophthalmology"},{"key":"2021122705113476300_R5","doi-asserted-by":"crossref","first-page":"1202","DOI":"10.1046\/j.1523-1747.2002.19526.x","article-title":"Common mutations in Arg304 of the p63 gene in ectrodactyly, ectodermal dysplasia, clefting syndrome: Lack of genotype-phenotype correlation and implications for mutation detection strategies","volume":"119","author":"Hamada","year":"2002","journal-title":"J Invest Dermatol"},{"key":"2021122705113476300_R6","doi-asserted-by":"crossref","first-page":"1957","DOI":"10.1002\/ajmg.a.35414","article-title":"A novel de novo missense mutation in TP63 underlying germline mosaicism in AEC syndrome: Implications for recurrence risk and prenatal diagnosis","volume":"158A","author":"Barbaro","year":"2012","journal-title":"Am J Med Genet A"},{"key":"2021122705113476300_R7","first-page":"87","article-title":"Gene therapy approaches for corneal diseases","author":"Di Iorio","year":"2013","journal-title":"New Insights on Biobanks"},{"key":"2021122705113476300_R8","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1016\/S0161-6420(00)00475-9","article-title":"Living related conjunctival limbal allograft for the treatment of stem cell 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epithelium","volume":"351","author":"Nishida","year":"2004","journal-title":"N Engl J Med"},{"key":"2021122705113476300_R12","doi-asserted-by":"crossref","first-page":"4660","DOI":"10.1167\/iovs.09-3377","article-title":"Persistence of transplanted oral mucosal epithelial cells in human cornea","volume":"50","author":"Chen","year":"2009","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"2021122705113476300_R13","doi-asserted-by":"crossref","first-page":"1325","DOI":"10.1167\/iovs.11-7744","article-title":"Cultured autologous oral mucosal epithelial cell sheet (CAOMECS) transplantation for the treatment of corneal limbal epithelial stem cell deficiency","volume":"53","author":"Burillon","year":"2012","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"2021122705113476300_R14","first-page":"2084","article-title":"Reconstruction of a human hemicornea through natural scaffolds compatible with the growth of corneal epithelial stem cells and stromal keratocytes","volume":"15","author":"Barbaro","year":"2009","journal-title":"Mol Vis"},{"key":"2021122705113476300_R15","doi-asserted-by":"crossref","first-page":"832","DOI":"10.1002\/gcc.22079","article-title":"Genetic instability of the tumor suppressor gene FHIT in normal human cells","volume":"52","author":"Palumbo","year":"2013","journal-title":"Genes Chromosomes Cancer"},{"key":"2021122705113476300_R16","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1007\/s00412-010-0279-4","article-title":"Replication dynamics at common fragile site FRA6E","volume":"119","author":"Palumbo","year":"2010","journal-title":"Chromosoma"},{"key":"2021122705113476300_R17","doi-asserted-by":"crossref","first-page":"926","DOI":"10.1136\/bjo.2009.164152","article-title":"Evaluation of ocular surface disorders: A new diagnostic tool based on impression cytology and confocal laser scanning microscopy","volume":"94","author":"Barbaro","year":"2010","journal-title":"Br J 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