Open Access Research

Congenital Dyserythropoietic Anemia Type II: molecular analysis and expression of the SEC23B Gene

Francesca Punzo12, Aida M Bertoli-Avella1, Saverio Scianguetta2, Fulvio Della Ragione3, Maddalena Casale2, Luisa Ronzoni4, Maria D Cappellini4, Gianluca Forni5, Ben A Oostra1 and Silverio Perrotta2*

Author Affiliations

1 Department of Clinical Genetics, Erasmus Medical Centre, Rotterdam, the Netherlands

2 Department of Paediatrics, Second University of Naples, Naples, Italy

3 Department of Biochemistry and Biophysics, "F. Cedrangolo," Second University of Naples, Naples, Italy

4 Department of Internal Medicine, Policlinic Foundation IRCCS, Milan University, Milan, Italy

5 Centro della Microcitemia e Anemie Congenite, Galliera Hospital, Genova, Italy

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Orphanet Journal of Rare Diseases 2011, 6:89  doi:10.1186/1750-1172-6-89

Published: 30 December 2011

Abstract

Background

Congenital dyserythropoietic anemia type II (CDAII), the most common form of CDA, is an autosomal recessive condition. CDAII diagnosis is based on invasive, expensive, and time consuming tests that are available only in specialized laboratories. The recent identification of SEC23B mutations as the cause of CDAII opens new possibilities for the molecular diagnosis of the disease. The aim of this study was to characterize molecular genomic SEC23B defects in 16 unrelated patients affected by CDAII and correlate the identified genetic alterations with SEC23B transcript and protein levels in erythroid precursors.

Methods

SEC23B was sequenced in 16 patients, their relatives and 100 control participants. SEC23B transcript level were studied by quantitative PCR (qPCR) in peripheral erythroid precursors and lymphocytes from the patients and healthy control participants. Sec23B protein content was analyzed by immunoblotting in samples of erythroblast cells from CDAII patients and healthy controls.

Results

All of the investigated cases carried SEC23B mutations on both alleles, with the exception of two patients in which a single heterozygous mutation was found. We identified 15 different SEC23B mutations, of which four represent novel mutations: p.Gln214Stop, p.Thr485Ala, p.Val637Gly, and p.Ser727Phe. The CDAII patients exhibited a 40-60% decrease of SEC23B mRNA levels in erythroid precursors when compared with the corresponding cell type from healthy participants. The largest decrease was observed in compound heterozygote patients with missense/nonsense mutations. In three patients, Sec23B protein levels were evaluated in erythroid precursors and found to be strictly correlated with the reduction observed at the transcript level. We also demonstrate that Sec23B mRNA expression levels in lymphocytes and erythroblasts are similar.

Conclusions

In this study, we identified four novel SEC23B mutations associated with CDAII disease. We also demonstrate that the genetic alteration results in a significant decrease of SEC23B transcript in erythroid precursors. Similar down-regulation was observed in peripheral lymphocytes, suggesting that the use of these cells might be sufficient in the identification of Sec23B gene alterations. Finally, we demonstrate that decreased Sec23B protein levels in erythroid precursors correlate with down-regulation of the SEC23B mRNA transcript.

Keywords:
Congenital dyserythropoietic anemia; CDA II; SEC23B; Red blood cell; Coat complex protein II