{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T13:07:31Z","timestamp":1792156051797,"version":"4.4.0"},"reference-count":28,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2022,1,22]],"date-time":"2022-01-22T00:00:00Z","timestamp":1642809600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Haemophilia"],"published-print":{"date-parts":[[2022,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>von Willebrand factor propeptide (VWFpp) plays an important role in VWF multimerization and storage. VWFpp mutations have been previously associated with types 1, 3 and 2A\/IIC von Willebrand disease (VWD).<\/jats:p><\/jats:sec><jats:sec><jats:title>Aims<\/jats:title><jats:p>To characterize the <jats:italic>novel<\/jats:italic> p.Thr274Pro variant identified in two unrelated type 1 VWD patients.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Phenotype tests were performed to evaluate patients\u2019 plasma and platelets following the current ISTH\u2010SSC guidelines. Molecular analysis was performed using next\u2010generation sequencing. The pcDNA3.1\u2010VWF\u2010WT and mutant pcDNA3.1\u2010VWF\u2010Thr274Pro expression vectors were transiently transfected into HEK293 cells to evaluate recombinant (r)VWF constitutive and regulated secretion. For the latter, the transfected cells were stimulated with phorbol\u201012\u2010myristate\u201013\u2010acetate. Immunofluorescence staining was performed to assess the localization of WT\u2010rVWF and Thr274Pro\u2010rVWF in endoplasmic reticulum, lysosomes, cis\u2010\/trans\u2010Golgi and pseudo\u2010Weibel Palade bodies.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Biochemical characterization of patients\u2019 plasma samples indicated a type 1 VWD diagnosis. Both patients were heterozygous for the p.Thr274Pro variant. Hybrid Thr274Pro\/WT\u2010rVWF showed a secretion reduction of 36\u00b14% according to patients\u2019 plasma VWF:Ag levels, whereas Thr274Pro\u2010rVWF secretion was strongly impaired (21\u00b12%). The amount of rVWF in cell lysates was nearly normal for both Thr274P (62\u00b117%) and Thr274Pro\/WT\u2010rVWF (72\u00b123%). The regulated secretion was impaired for Thr274Pro\/WT\u2010rVWF, whereas Thr274Pro\u2010rVWF was not released at all. Immunofluorescence staining revealed no particular differences between WT and Thr274Pro\u2010rVWF, although Thr274Pro\u2010rVWF showed less pseudo\u2010Weibel Palade bodies with a rounder shape than WT\u2010rVWF.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The <jats:italic>novel<\/jats:italic> p.Thr274Pro mutation has a dominant effect and it is responsible of patients\u2019 type 1 VWD phenotype through a combined mechanism of reduced synthesis, impaired secretion and multimerization.<\/jats:p><\/jats:sec>","DOI":"10.1111\/hae.14494","type":"journal-article","created":{"date-parts":[[2022,1,22]],"date-time":"2022-01-22T10:24:49Z","timestamp":1642847089000},"page":"292-300","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["The dominant p.Thr274Pro mutation in the von Willebrand factor propeptide causes the von Willebrand disease type 1 phenotype in two unrelated patients"],"prefix":"10.1111","volume":"28","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0778-2271","authenticated-orcid":false,"given":"Maria Teresa","family":"Pagliari","sequence":"first","affiliation":[{"name":"Fondazione IRCCS Ca\u2019 Granda Ospedale Maggiore Policlinico, Angelo Bianchi Bonomi Hemophilia and Thrombosis Center and Universit\u00e0 degli Studi di Milano, Department of Pathophysiology and Transplantation  Milan Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luciano","family":"Baronciani","sequence":"additional","affiliation":[{"name":"Fondazione IRCCS Ca\u2019 Granda Ospedale Maggiore Policlinico Angelo Bianchi Bonomi Hemophilia and Thrombosis Center  Milan Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2013-0043","authenticated-orcid":false,"given":"Chiara","family":"Cordiglieri","sequence":"additional","affiliation":[{"name":"National Institute of Molecular Genetics \u2018Romeo e Enrica Invernizzi\u2019 \u2010 INGM  Milan Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paola","family":"Colpani","sequence":"additional","affiliation":[{"name":"Fondazione IRCCS Ca\u2019 Granda Ospedale Maggiore Policlinico Angelo Bianchi Bonomi Hemophilia and Thrombosis Center  Milan Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giovanna","family":"Cozzi","sequence":"additional","affiliation":[{"name":"Fondazione IRCCS Ca\u2019 Granda Ospedale Maggiore Policlinico Angelo Bianchi Bonomi Hemophilia and Thrombosis Center  Milan Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8456-7519","authenticated-orcid":false,"given":"Simona M.","family":"Siboni","sequence":"additional","affiliation":[{"name":"Fondazione IRCCS Ca\u2019 Granda Ospedale Maggiore Policlinico Angelo Bianchi Bonomi Hemophilia and Thrombosis Center  Milan 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