{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,5]],"date-time":"2026-09-05T03:52:39Z","timestamp":1788580359857,"version":"build-2803163510"},"reference-count":43,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2020,5,22]],"date-time":"2020-05-22T00:00:00Z","timestamp":1590105600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Acad Dermatol Venereol"],"published-print":{"date-parts":[[2020,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>A lack or dysfunction of the anchoring protein laminin\u2010332 in the basement membrane leads to the skin blistering disorder junctional epidermolysis bullosa (JEB). The mutation c.628G&gt;A in the gene <jats:italic>LAMB3<\/jats:italic> encoding the laminin \u03b23\u2010chain is associated with generalized intermediate JEB; it may introduce an amino acid substitution (p.Glu210Lys) or disrupt splicing.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objective<\/jats:title><jats:p>This retrospective study aimed at determining the effects of aberrant splicing on the JEB phenotype.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p><jats:italic>LAMB3<\/jats:italic> transcription was analysed in two siblings compound heterozygous for the <jats:italic>LAMB3<\/jats:italic> mutations p.Glu210Lys and p.Arg635* with a diverging JEB phenotype from late childhood on. Laminin\u2010332 levels in skin sections and in cultured keratinocytes were investigated by immunofluorescence staining. Real\u2010time PCR was used to quantify <jats:italic>LAMB3<\/jats:italic> expression in keratinocytes. RNA splice variants were identified by subcloning of a <jats:italic>LAMB3<\/jats:italic> cDNA fraction and subsequent DNA sequencing. Structural models of laminin\u2010332 helped to assess the impact of certain mutations on laminin\u2010332 folding.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Both siblings showed diminished <jats:italic>LAMB3<\/jats:italic> expression. Laminin\u2010332 was equally reduced in skin sections obtained during infancy but differed in keratinocytes isolated during adolescence. Although aberrant <jats:italic>LAMB3<\/jats:italic> splicing with 26 variants was detected in both patients, splicing differed significantly: the full\u2010length <jats:italic>LAMB3<\/jats:italic> transcript harbouring the p.Glu210Lys mutation was found more often in the patient affected less severely (14\/108 vs. 5\/106 clones; <jats:italic>P<\/jats:italic>\u00a0=\u00a00.03). Structural modelling predicted that several deletions in <jats:italic>LAMB3<\/jats:italic>, but not the point mutation p.Glu210Lys, have an effect on laminin\u2010332 folding and secretion.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Differential <jats:italic>LAMB3<\/jats:italic> mRNA splicing in the patients may explain the disparate JEB phenotype. By elucidating the regulation of laminin\u2010332 gene expression, these findings may contribute to the development of therapeutic strategies for JEB and might help to understand phenotype modification by splice\u2010site mutations in other hereditary diseases.<\/jats:p><\/jats:sec>","DOI":"10.1111\/jdv.16332","type":"journal-article","created":{"date-parts":[[2020,3,3]],"date-time":"2020-03-03T10:03:21Z","timestamp":1583229801000},"page":"2127-2134","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Aberrant splicing as potential modifier of the phenotype of junctional epidermolysis bullosa"],"prefix":"10.1111","volume":"34","author":[{"given":"R.","family":"Mittwollen","sequence":"first","affiliation":[{"name":"Department of Pediatrics University Hospital Erlangen Erlangen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S.","family":"Wohlfart","sequence":"additional","affiliation":[{"name":"Department of Pediatrics University Hospital Erlangen Erlangen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J.","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Pediatrics University Hospital Erlangen Erlangen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"E.","family":"Grosch","sequence":"additional","affiliation":[{"name":"Department of Dermatology University Hospital Erlangen Erlangen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6066-507X","authenticated-orcid":false,"given":"C.","family":"Has","sequence":"additional","affiliation":[{"name":"Department of Dermatology University Medical Center Freiburg Freiburg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"E.","family":"Hohenester","sequence":"additional","affiliation":[{"name":"Department of Life Sciences Imperial College London London UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"H.","family":"Schneider","sequence":"additional","affiliation":[{"name":"Department of Pediatrics University Hospital Erlangen Erlangen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8821-5775","authenticated-orcid":false,"given":"J.","family":"Hammersen","sequence":"additional","affiliation":[{"name":"Department of Pediatrics University Hospital Erlangen Erlangen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2020,5,22]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00439-001-0630-1"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1067\/mjd.2002.119673"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejcb.2006.07.004"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.4161\/cam.22418"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaad.2014.01.903"},{"key":"e_1_2_6_7_1","first-page":"849","article-title":"Junctional epidermolysis bullosa with LAMB3 splice\u2010site mutations","volume":"95","author":"Kiritsi D","year":"2015","journal-title":"Acta Derm Venereol"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1007\/s13353-018-0444-7"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jdermsci.2005.05.006"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00439-017-1789-4"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-0625.1998.tb00309.x"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/labinvest.3700164"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.0022-202X.2004.23302.x"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1136\/jmg.2005.039685"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1111\/1523-1747.ep12605904"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1111\/1523-1747.ep12340210"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2133.1998.02377.x"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI30465"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.1998.8180"},{"key":"e_1_2_6_20_1","first-page":"859","article-title":"LAMB3 mutations in generalized atrophic benign epidermolysis bullosa: consequences at the mRNA and protein levels","volume":"78","author":"Pulkkinen L","year":"1998","journal-title":"Lab Invest"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/jid.2014.466"},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.gt.3302832"},{"key":"e_1_2_6_23_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0042473"},{"key":"e_1_2_6_24_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.187.8.1273"},{"key":"e_1_2_6_25_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M402455200"},{"key":"e_1_2_6_26_1","doi-asserted-by":"publisher","DOI":"10.4161\/cam.23132"},{"key":"e_1_2_6_27_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/15.17.7155"},{"key":"e_1_2_6_28_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMc0809896"},{"key":"e_1_2_6_29_1","doi-asserted-by":"publisher","DOI":"10.1111\/bjd.14374"},{"key":"e_1_2_6_30_1","doi-asserted-by":"crossref","first-page":"e0192994","DOI":"10.1371\/journal.pone.0192994","article-title":"A \"late\u2010but\u2010fitter revertant cell\" explains the high frequency of revertant mosaicism in epidermolysis bullosa","volume":"13","author":"Akker PC","year":"2018","journal-title":"PLoS One"},{"key":"e_1_2_6_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0960-9822(00)80056-1"},{"key":"e_1_2_6_32_1","doi-asserted-by":"publisher","DOI":"10.2340\/00015555-2439"},{"key":"e_1_2_6_33_1","doi-asserted-by":"publisher","DOI":"10.1111\/bjd.15690"},{"key":"e_1_2_6_34_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.1004068"},{"key":"e_1_2_6_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.matbio.2018.10.003"},{"key":"e_1_2_6_36_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2133.1999.03170.x"},{"key":"e_1_2_6_37_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0071143"},{"key":"e_1_2_6_38_1","doi-asserted-by":"publisher","DOI":"10.12688\/f1000research.5654.1"},{"key":"e_1_2_6_39_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrg2415"},{"key":"e_1_2_6_40_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms3771"},{"key":"e_1_2_6_41_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1523-1747.1999.00709.x"},{"key":"e_1_2_6_42_1","doi-asserted-by":"publisher","DOI":"10.1093\/hmg\/10.21.2453"},{"key":"e_1_2_6_43_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-8651-4_2"},{"key":"e_1_2_6_44_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature24487"}],"container-title":["Journal of the European Academy of Dermatology and Venereology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fjdv.16332","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/jdv.16332","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/jdv.16332","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/jdv.16332","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,3]],"date-time":"2023-09-03T02:25:01Z","timestamp":1693707901000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/jdv.16332"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,5,22]]},"references-count":43,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2020,9]]}},"alternative-id":["10.1111\/jdv.16332"],"URL":"https:\/\/doi.org\/10.1111\/jdv.16332","archive":["Portico"],"relation":{},"ISSN":["0926-9959","1468-3083"],"issn-type":[{"value":"0926-9959","type":"print"},{"value":"1468-3083","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,5,22]]},"assertion":[{"value":"2019-09-22","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-02-06","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-05-22","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}