Congenital dyserythropoietic anemia type II: Molecular analysis and expression of the SEC23B Gene

Francesca Punzo, Aida M. Bertoli-Avella, Saverio Scianguetta, Fulvio Della Ragione, Maddalena Casale, Luisa Ronzoni, Maria D. Cappellini, Gianluca Forni, Ben A. Oostra, Silverio Perrotta

Research output: Contribution to journalArticle

14 Citations (Scopus)

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.

Original languageEnglish
Article number89
JournalOrphanet Journal of Rare Diseases
Volume6
Issue number1
DOIs
Publication statusPublished - 2011

Fingerprint

Congenital Dyserythropoietic Anemia
Gene Expression
Mutation
Erythroblasts
Lymphocytes
Messenger RNA
Healthy Volunteers
Proteins
Down-Regulation
Nonsense Codon
Missense Mutation
Heterozygote
Immunoblotting
Alleles

Keywords

  • CDA II
  • Coat complex protein II
  • Congenital dyserythropoietic anemia
  • Red blood cell
  • SEC23B

ASJC Scopus subject areas

  • Medicine(all)
  • Genetics(clinical)
  • Pharmacology (medical)

Cite this

Punzo, F., Bertoli-Avella, A. M., Scianguetta, S., Della Ragione, F., Casale, M., Ronzoni, L., ... Perrotta, S. (2011). Congenital dyserythropoietic anemia type II: Molecular analysis and expression of the SEC23B Gene. Orphanet Journal of Rare Diseases, 6(1), [89]. https://doi.org/10.1186/1750-1172-6-89

Congenital dyserythropoietic anemia type II : Molecular analysis and expression of the SEC23B Gene. / Punzo, Francesca; Bertoli-Avella, Aida M.; Scianguetta, Saverio; Della Ragione, Fulvio; Casale, Maddalena; Ronzoni, Luisa; Cappellini, Maria D.; Forni, Gianluca; Oostra, Ben A.; Perrotta, Silverio.

In: Orphanet Journal of Rare Diseases, Vol. 6, No. 1, 89, 2011.

Research output: Contribution to journalArticle

Punzo, F, Bertoli-Avella, AM, Scianguetta, S, Della Ragione, F, Casale, M, Ronzoni, L, Cappellini, MD, Forni, G, Oostra, BA & Perrotta, S 2011, 'Congenital dyserythropoietic anemia type II: Molecular analysis and expression of the SEC23B Gene', Orphanet Journal of Rare Diseases, vol. 6, no. 1, 89. https://doi.org/10.1186/1750-1172-6-89
Punzo, Francesca ; Bertoli-Avella, Aida M. ; Scianguetta, Saverio ; Della Ragione, Fulvio ; Casale, Maddalena ; Ronzoni, Luisa ; Cappellini, Maria D. ; Forni, Gianluca ; Oostra, Ben A. ; Perrotta, Silverio. / Congenital dyserythropoietic anemia type II : Molecular analysis and expression of the SEC23B Gene. In: Orphanet Journal of Rare Diseases. 2011 ; Vol. 6, No. 1.
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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.",
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author = "Francesca Punzo and Bertoli-Avella, {Aida M.} and Saverio Scianguetta and {Della Ragione}, Fulvio and Maddalena Casale and Luisa Ronzoni and Cappellini, {Maria D.} and Gianluca Forni and Oostra, {Ben A.} and Silverio Perrotta",
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T2 - Molecular analysis and expression of the SEC23B Gene

AU - Punzo, Francesca

AU - Bertoli-Avella, Aida M.

AU - Scianguetta, Saverio

AU - Della Ragione, Fulvio

AU - Casale, Maddalena

AU - Ronzoni, Luisa

AU - Cappellini, Maria D.

AU - Forni, Gianluca

AU - Oostra, Ben A.

AU - Perrotta, Silverio

PY - 2011

Y1 - 2011

N2 - 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.

AB - 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.

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KW - Coat complex protein II

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KW - Red blood cell

KW - SEC23B

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