Congenital dyserythropoietic anemia type II: molecular analysis and expression of the SEC23B gene.
Congenital dyserythropoietic anemia type II: molecular analysis and expression of the SEC23B gene.
复制标题
先天性颗粒嗜性贫血II型：Sec23b基因的分子分析和表达。
DOI：
10.1186/1750-1172-6-89
复制
发表时间：
2011-12-30
影响因子：
3.7
通讯作者：
Perrotta S
中科院分区：
文献类型：
--
作者：
Punzo F;Bertoli-Avella AM;Scianguetta S;Della Ragione F;Casale M;Ronzoni L;Cappellini MD;Forni G;Oostra BA;Perrotta S
先天性红细胞生成性贫血II型(CDAII)是CDA最常见的形式，是一种常染色体隐性遗传性疾病。CDAII诊断基于侵入性、昂贵和耗时的测试，这些测试只有在专门的实验室才能获得。最近发现的导致CDAII的SEC23B突变为CDAII的分子诊断开辟了新的可能性。本研究的目的是分析16例非血缘关系的CDAII患者的SEC23B基因缺陷，并将发现的基因改变与红系前体细胞的SEC23B转录本和蛋白水平相关联。对16名患者及其亲属和100名对照受试者进行了SEC23B基因测序。采用定量聚合酶链式反应(QPCR)检测患者和健康对照组外周血红系前体细胞和淋巴细胞中SEC23B转录水平。免疫印迹法检测CDAII患者和健康对照组红细胞中Sec23B蛋白含量。除两例患者发现单一杂合性突变外，其余病例均携带SEC23B基因突变。我们鉴定了15个不同的SEC23B突变，其中4个代表新的突变：p.Gln214Stop，p.Thr485Ala，p.Val637Gly和p.Ser727Phe。与健康受试者的相应细胞类型相比，CDAII患者红系前体细胞中SEC23B基因表达水平降低了40-60%。在错义/无义突变的复合杂合子患者中，下降幅度最大。在三名患者中，对红系前体细胞中Sec23B蛋白水平进行了评估，发现其与转录本水平上观察到的减少密切相关。我们还证明了Sec23B在淋巴细胞和红细胞中的表达水平是相似的。在这项研究中，我们发现了四个与CDAII病相关的新的SEC23B突变。我们还证明了基因改变导致红系前体细胞中SEC23B转录本的显著减少。在外周血淋巴细胞中也观察到类似的下调，提示利用这些细胞可能足以鉴定Sec23B基因的改变。最后，我们证明了红系前体细胞中Sec23B蛋白水平的降低与SEC23B基因转录的下调有关。
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. 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. 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. 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.
登录
查看更多内容
影响因子：
6.5
作者：
CROOKSTON, JH;CROOKSTON, MC;LEWIS, SM
通讯作者：
LEWIS, SM
影响因子：
2.4
作者：
Amir, Achiya;Dgany, Orly;Tamary, Hannah
通讯作者：
Tamary, Hannah
影响因子：
30.8
作者：
Schwarz, Klaus;Iolascon, Achille;Heimpel, Hermann
通讯作者：
Heimpel, Hermann
影响因子：
21.3
作者：
Antonny, B;Madden, D;Schekman, R
通讯作者：
Schekman, R
影响因子：
2.3
作者：
Fermo, Elisa;Bianchi, Paola;Zanella, Alberto
通讯作者：
Zanella, Alberto