Sofia Vatsiou1, Maria Zamanakou2, Gedeon Loules2, Fotis Psarros3, Faidra Parsopoulou1, Dorottya Csuka4, Anna Valerieva5, Maria Staevska5, Grzegorz Porebski6, Krystyna Obtulowicz6, Markus Magerl7, Marcus Maurer7, Matthaios Speletas8, Henriette Farkas4, Anastasios E Germenis9. 1. CeMIA SA, Larissa, Greece; Department of Immunology & Histocompatibility, University of Thessaly, School of Health Sciences, Faculty of Medicine, Larissa, Greece. 2. CeMIA SA, Larissa, Greece. 3. Department of Allergology, Navy Hospital, Athens, Greece. 4. Hungarian Angioedema Reference Center, 3rd Department of Internal Medicine, Semmelweis University, Budapest, Hungary. 5. Clinic of Allergy and Asthma, Medical University of Sofia, Sofia, Bulgaria. 6. Department of Clinical and Environmental Allergology, Jagiellonian University Medical College, Krakow, Poland. 7. Department of Dermatology and Allergy, Charité - Universitätsmedizin Berlin, Berlin, Germany. 8. Department of Immunology & Histocompatibility, University of Thessaly, School of Health Sciences, Faculty of Medicine, Larissa, Greece. 9. CeMIA SA, Larissa, Greece; Department of Immunology & Histocompatibility, University of Thessaly, School of Health Sciences, Faculty of Medicine, Larissa, Greece. Electronic address: agermen@med.uth.gr.
Abstract
BACKGROUND: In about 5% of patients with hereditary angioedema due to C1-inhibitor deficiency (C1-INH-HAE) no mutation in the SERPING1 gene is detected. METHODS: C1-INH-HAE cases with no mutation in the coding region of SERPING1 after conventional genotyping were examined for defects in the intronic or untranslated regions of the gene. Using a next-generation sequencing (NGS) platform targeting the entire SERPING1, 14 unrelated C1-INH-HAE patients with no detectable mutations in the coding region of the gene were sequenced. Detected variants with a global minor allele frequency lower than the frequency of C1-INH-HAE (0.002%), were submitted to in silico analysis using ten different bioinformatics tools. Pedigree analysis and examination of their pathogenic effect on the RNA level were performed for filtered in variants. RESULTS: In two unrelated patients, the novel mutation c.-22-155G > T was detected in intron 1 of the SERPING1 gene by the use NGS and confirmed by Sanger sequencing. All bioinformatics tools predicted that the variant causes a deleterious effect on the gene and pedigree analysis showed its co-segregation with the disease. Degradation of the mutated allele was demonstrated by the loss of heterozygosity on the cDNA level. According to the American College of Medical Genetics and Genomics 2015 guidelines the c.-22-155G > T was curated as pathogenic. CONCLUSIONS: For the first time, a deep intronic mutation that was detected by NGS in the SERPING1 gene, was proven pathogenic for C1-INH-HAE. Therefore, advanced DNA sequencing methods should be performed in cases of C1-INH-HAE where standard approaches fail to uncover the genetic alteration.
BACKGROUND: In about 5% of patients with hereditary angioedema due to C1-inhibitor deficiency (C1-INH-HAE) no mutation in the SERPING1 gene is detected. METHODS:C1-INH-HAE cases with no mutation in the coding region of SERPING1 after conventional genotyping were examined for defects in the intronic or untranslated regions of the gene. Using a next-generation sequencing (NGS) platform targeting the entire SERPING1, 14 unrelated C1-INH-HAEpatients with no detectable mutations in the coding region of the gene were sequenced. Detected variants with a global minor allele frequency lower than the frequency of C1-INH-HAE (0.002%), were submitted to in silico analysis using ten different bioinformatics tools. Pedigree analysis and examination of their pathogenic effect on the RNA level were performed for filtered in variants. RESULTS: In two unrelated patients, the novel mutation c.-22-155G > T was detected in intron 1 of the SERPING1 gene by the use NGS and confirmed by Sanger sequencing. All bioinformatics tools predicted that the variant causes a deleterious effect on the gene and pedigree analysis showed its co-segregation with the disease. Degradation of the mutated allele was demonstrated by the loss of heterozygosity on the cDNA level. According to the American College of Medical Genetics and Genomics 2015 guidelines the c.-22-155G > T was curated as pathogenic. CONCLUSIONS: For the first time, a deep intronic mutation that was detected by NGS in the SERPING1 gene, was proven pathogenic for C1-INH-HAE. Therefore, advanced DNA sequencing methods should be performed in cases of C1-INH-HAE where standard approaches fail to uncover the genetic alteration.
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Authors: Carina M Mathey; Carlo Maj; Annika B Scheer; Julia Fazaal; Bettina Wedi; Dorothea Wieczorek; Philipp M Amann; Harald Löffler; Lukas Koch; Clemens Schöffl; Heinrich Dickel; Nomun Ganjuur; Thorsten Hornung; Susann Forkel; Jens Greve; Gerda Wurpts; Pär Hallberg; Anette Bygum; Christian Von Buchwald; Malgorzata Karawajczyk; Michael Steffens; Julia Stingl; Per Hoffmann; Stefanie Heilmann-Heimbach; Elisabeth Mangold; Kerstin U Ludwig; Eva R Rasmussen; Mia Wadelius; Bernhardt Sachs; Markus M Nöthen; Andreas J Forstner Journal: Front Genet Date: 2022-07-18 Impact factor: 4.772