Identification of a novel variant (c.1-111A>G) located in GATA-1 motif of RHCE proximal promoter in two Chinese patients with the rare D-- phenotype.

IF 2.5 3区 医学 Q2 HEMATOLOGY
Transfusion Pub Date : 2025-03-23 DOI:10.1111/trf.18223
Ran Zhang, Jizhi Wen, Changjiu Tang, Shuangshuang Jia, Qi Chen, Yanli Ji
{"title":"Identification of a novel variant (c.1-111A>G) located in GATA-1 motif of RHCE proximal promoter in two Chinese patients with the rare D-- phenotype.","authors":"Ran Zhang, Jizhi Wen, Changjiu Tang, Shuangshuang Jia, Qi Chen, Yanli Ji","doi":"10.1111/trf.18223","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>D-- is a rare phenotype lacking the expression of the C, c, E, and e antigens and several high-frequency antigens on the red cells. Anti-Rh17 (Hr0) could be developed in individuals with the D-- phenotype to cause hemolytic transfusion reactions (HTR) and hemolytic disease of the fetus and newborn (HDFN). Nuleotide(s) change of the RHCE gene and RHCE-D-CE hybrid alleles are the common molecular basis of the D-- phenotype.</p><p><strong>Study design and methods: </strong>One D-- Chinese patient detected in routine RhD and RhCE serologic testing and another D-- Chinese patient identified with anti-Rh17 were recruited. Further RHD, RHCE, and RHAG whole gene sequences were analyzed using the PacBio sequencing. A dual-luciferase reporter assay was performed to verify the effect of the variant identified in the promoter of the RHCE gene on the transcriptional activity of the reporter gene in vitro.</p><p><strong>Results: </strong>A homozygous RHCE*Ce(1-111G)/Ce(1-111G) genotype and a heterozygous RHCE*CeN.08/Ce(1-111G) genotype carried one novel variant (c.1-111A>G) located in the GATA-1 motif of the RHCE proximal promoter was identified in two D-- patients, respectively. In the reporter assay, the luciferase transcriptional activity of the mutant RHCE promoter [c.1-111G] construct reduced from ~1.0 to 0.28 relative luciferase activity normalized to RHCE wild-type, with a ~72% reduction rate.</p><p><strong>Conclusion: </strong>The novel variant of the GATA-1 motif of the RHCE proximal promoter was identified to diminish the binding of the GATA-1 transcription factor and markedly down-regulate the transcription activity of the RHCE gene to abolish the expression of RhCE antigens, causing the rare D-- phenotype.</p>","PeriodicalId":23266,"journal":{"name":"Transfusion","volume":" ","pages":""},"PeriodicalIF":2.5000,"publicationDate":"2025-03-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Transfusion","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1111/trf.18223","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"HEMATOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

Background: D-- is a rare phenotype lacking the expression of the C, c, E, and e antigens and several high-frequency antigens on the red cells. Anti-Rh17 (Hr0) could be developed in individuals with the D-- phenotype to cause hemolytic transfusion reactions (HTR) and hemolytic disease of the fetus and newborn (HDFN). Nuleotide(s) change of the RHCE gene and RHCE-D-CE hybrid alleles are the common molecular basis of the D-- phenotype.

Study design and methods: One D-- Chinese patient detected in routine RhD and RhCE serologic testing and another D-- Chinese patient identified with anti-Rh17 were recruited. Further RHD, RHCE, and RHAG whole gene sequences were analyzed using the PacBio sequencing. A dual-luciferase reporter assay was performed to verify the effect of the variant identified in the promoter of the RHCE gene on the transcriptional activity of the reporter gene in vitro.

Results: A homozygous RHCE*Ce(1-111G)/Ce(1-111G) genotype and a heterozygous RHCE*CeN.08/Ce(1-111G) genotype carried one novel variant (c.1-111A>G) located in the GATA-1 motif of the RHCE proximal promoter was identified in two D-- patients, respectively. In the reporter assay, the luciferase transcriptional activity of the mutant RHCE promoter [c.1-111G] construct reduced from ~1.0 to 0.28 relative luciferase activity normalized to RHCE wild-type, with a ~72% reduction rate.

Conclusion: The novel variant of the GATA-1 motif of the RHCE proximal promoter was identified to diminish the binding of the GATA-1 transcription factor and markedly down-regulate the transcription activity of the RHCE gene to abolish the expression of RhCE antigens, causing the rare D-- phenotype.

在两例罕见的D-表型的中国患者中鉴定了一个位于RHCE近端启动子GATA-1基序的新变异(c.1-111A>G)。
背景:D-是一种罕见的表型,缺乏C, C, E和E抗原以及红细胞上的几种高频抗原的表达。抗rh17 (Hr0)可在具有D-表型的个体中发展,引起溶血性输血反应(HTR)和胎儿和新生儿溶血性疾病(hddn)。RHCE基因和RHCE-D- ce杂交等位基因的核苷酸变化是D-表型的共同分子基础。研究设计和方法:招募1例常规RhD和RhCE血清学检测中检测到的D-中国患者和另1例抗rh17检测出的D-中国患者。采用PacBio测序进一步分析RHD、RHCE和RHAG全基因序列。双荧光素酶报告基因实验验证了在RHCE基因启动子中发现的变异对报告基因体外转录活性的影响。结果:在2例D-患者中分别鉴定出纯合子RHCE*Ce(1-111G)/Ce(1-111G)基因型和杂合子RHCE*CeN.08/Ce(1-111G)基因型携带一个位于RHCE近端启动子GATA-1基序的新变体(c.1-111A>G)。在报告基因实验中,突变体RHCE启动子的荧光素酶转录活性[c]。1-111G]构建体的相对荧光素酶活性从~1.0降低到0.28,还原率为~72%。结论:发现了RHCE近端启动子GATA-1基序的新变异,该变异减少了GATA-1转录因子的结合,显著下调了RHCE基因的转录活性,从而消除了RHCE抗原的表达,导致了罕见的D-表型。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Transfusion
Transfusion 医学-血液学
CiteScore
4.70
自引率
20.70%
发文量
426
审稿时长
1 months
期刊介绍: TRANSFUSION is the foremost publication in the world for new information regarding transfusion medicine. Written by and for members of AABB and other health-care workers, TRANSFUSION reports on the latest technical advances, discusses opposing viewpoints regarding controversial issues, and presents key conference proceedings. In addition to blood banking and transfusion medicine topics, TRANSFUSION presents submissions concerning patient blood management, tissue transplantation and hematopoietic, cellular, and gene therapies.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信