A functional assay for the clinical annotation of genetic variants of uncertain significance in Diamond-Blackfan anemia
Open Access
- 28 May 2018
- journal article
- research article
- Published by Hindawi Limited in Human Mutation
- Vol. 39 (8), 1102-1111
- https://doi.org/10.1002/humu.23551
Abstract
Diamond‐Blackfan anemia (DBA) is a rare genetic hypoplasia of erythroid progenitors characterized by mild to severe anemia and associated with congenital malformations. Clinical manifestations in DBA patients are quite variable and genetic testing has become a critical factor in establishing a diagnosis of DBA. The majority of DBA cases are due to heterozygous loss‐of‐function mutations in ribosomal protein (RP) genes. Causative mutations are fairly straightforward to identify in the case of large deletions and frameshift and nonsense mutations found early in a protein coding sequence, but diagnosis becomes more challenging in the case of missense mutations and small in‐frame indels. Our group recently characterized the phenotype of lymphoblastoid cell lines established from DBA patients with pathogenic lesions in RPS19 and observed that defective pre‐rRNA processing, a hallmark of the disease, was rescued by lentiviral vectors expressing wild‐type RPS19. Here, we use this complementation assay to determine whether RPS19 variants of unknown significance are capable of rescuing pre‐rRNA processing defects in these lymphoblastoid cells as a means of assessing the effects of these sequence changes on the function of the RPS19 protein. This approach will be useful in differentiating pathogenic mutations from benign polymorphisms in identifying causative genes in DBA patients. This article is protected by copyright. All rights reservedKeywords
Funding Information
- Diamond Blackfan Anemia Foundation
- Fondazione Europea per la DBA
- Gruppo di Sostegno DBA Italia
- Banca del Piemonte
- E-RARE EuroDBA Consortium (BMBF #01GM1301, 01GM1609)
- European Research Council (261178)
- Fondazione Telethon (GGP13177)
This publication has 63 references indexed in Scilit:
- Exome sequencing identifies GATA1 mutations resulting in Diamond-Blackfan anemiaJCI Insight, 2012
- Guidelines for splicing analysis in molecular diagnosis derived from a set of 327 combined in silico/in vitro studies on BRCA1 and BRCA2 variantsHuman Mutation, 2012
- Ribosomal Protein Genes RPS10 and RPS26 Are Commonly Mutated in Diamond-Blackfan AnemiaAmerican Journal of Human Genetics, 2010
- Posttranscriptional down-regulation of small ribosomal subunit proteins correlates with reduction of 18S rRNA in RPS19 deficiencyFEBS Letters, 2009
- Diamond-Blackfan Anemia: Diagnosis, Treatment, and Molecular PathogenesisHematology/Oncology Clinics of North America, 2009
- Ribosomal Protein L5 and L11 Mutations Are Associated with Cleft Palate and Abnormal Thumbs in Diamond-Blackfan Anemia PatientsAmerican Journal of Human Genetics, 2008
- Diagnosing and treating Diamond Blackfan anaemia: results of an international clinical consensus conferenceBritish Journal of Haematology, 2008
- Molecular basis of Diamond Blackfan anemia: structure and function analysis of RPS19Nucleic Acids Research, 2007
- Ribosomal Protein S24 Gene Is Mutated in Diamond-Blackfan AnemiaAmerican Journal of Human Genetics, 2006
- Human RPS19, the gene mutated in Diamond-Blackfan anemia, encodes a ribosomal protein required for the maturation of 40S ribosomal subunitsBlood, 2006