Diagnosis of vascular Ehlers-Danlos syndrome in Italy: clinical findings and novel COL3A1 mutations.

In this study, we describe the clinical features, the behaviour of dermal fibroblasts type III collagen (COLLIII) and COL3A1 gene mutations in 22 Italian vEDS patients. All subjects provided written, informed consent and authorised the processing of their personal data according to Italian bioethics laws. Control and patient skin fibroblasts were established from skin biopsies and were metabolically labelled. Pepsin-purified collagens (COLLs) from the culture medium and cell layer were run in a 6% SDS/PAGE, as previously described [9]. COLLIII from the medium was evaluated based on the a1(III)3/a1(I) + a2(I) COLLs chains ratio; the control ratio range (0.09–0.15) was established by analysis of 10 control fibroblast strains. Direct sequencing of cDNA and/or gDNA was used to search for mutations in the COL3A1 gene by standard procedures. All of the mutations identified by cDNA analysis were verified by gDNA sequencing and were investigated in the patients’ parents; novel mutations were confirmed by analysing the alleles of 400 healthy donors.