A common TBP-binding site mutation elevates fetal hemoglobin levels by competitive globin switching change in β-thalassemia
Abstract: β-Thalassemia is a common monogenic disorder caused by genetic defects in β-globin genes (HBB) resulting in imbalanced synthesis of α-/β-globin and ineffective erythropoiesis. It has been well documented that patients with β-thalassemia, or even carriers, mostly experience reactivation of...
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Elsevier
2025-07-01
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| Series: | Blood Advances |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2473952925002095 |
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| author | Mengyang Song Xiaolei Wei Hualei Luo Jueheng Wang Yuhua Ye Lang Qin Chao Niu Yong Long Xingmin Wang Congwen Shao Miao Yu Feng Gu Xinhua Zhang Xiangmin Xu |
| author_facet | Mengyang Song Xiaolei Wei Hualei Luo Jueheng Wang Yuhua Ye Lang Qin Chao Niu Yong Long Xingmin Wang Congwen Shao Miao Yu Feng Gu Xinhua Zhang Xiangmin Xu |
| author_sort | Mengyang Song |
| collection | DOAJ |
| description | Abstract: β-Thalassemia is a common monogenic disorder caused by genetic defects in β-globin genes (HBB) resulting in imbalanced synthesis of α-/β-globin and ineffective erythropoiesis. It has been well documented that patients with β-thalassemia, or even carriers, mostly experience reactivation of fetal hemoglobin (Hb F), but its underlying mechanisms are incompletely understood. We took advantage of a previously established cohort of 1142 patients with β-thalassemia with diverse thalassemic mutations subjected to targeted next-generation sequencing. Genotype-phenotype association studies demonstrated that the HBB:c.-78A>G had a remarkable effect on the elevation of Hb F levels compared with other β-thalassemic mutations. To experimentally validate this conclusion, the ribonucleoprotein transfection complex through homology-directed repair by electroporation was performed, from which we observed a consistent increase of Hb F expression in both HUDEP-2 and primary CD34+ cell lines. Furthermore, chromatin immunoprecipitation–quantitative polymerase chain reaction, dual-luciferase reporter assay, and circular chromosome conformation capture (4C) assays validated a decreased occupancy of the HBB TATA box by TATA-binding protein (TBP), leading to boosted expression of γ-globin genes by enhanced interaction between locus control regions (LCRs) and γ-globin gene promoters. The patient-based investigation and experimental validations presented in this study might lead to a better understanding of stage-specific globin-gene expression mediated by competitive binding of distal enhancers (LCRs). |
| format | Article |
| id | doaj-art-27c42ffdc7794057a74cf6cbcbc4c15f |
| institution | OA Journals |
| issn | 2473-9529 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Blood Advances |
| spelling | doaj-art-27c42ffdc7794057a74cf6cbcbc4c15f2025-08-20T02:20:51ZengElsevierBlood Advances2473-95292025-07-019133159316910.1182/bloodadvances.2024013695A common TBP-binding site mutation elevates fetal hemoglobin levels by competitive globin switching change in β-thalassemiaMengyang Song0Xiaolei Wei1Hualei Luo2Jueheng Wang3Yuhua Ye4Lang Qin5Chao Niu6Yong Long7Xingmin Wang8Congwen Shao9Miao Yu10Feng Gu11Xinhua Zhang12Xiangmin Xu13Department of Innovation Center for Diagnostics and Treatment of Thalassemia, Nanfang Hospital, Southern Medical University, Guangzhou, China; Department of Medical Genetics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, ChinaDepartment of Hematology, Nanfang Hospital, Southern Medical University, Guangzhou, China; Department of Clinical Medical Research Center of Hematological Diseases of Guangdong Province, Guangzhou, ChinaDepartment of Innovation Center for Diagnostics and Treatment of Thalassemia, Nanfang Hospital, Southern Medical University, Guangzhou, China; Department of Medical Genetics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, ChinaDepartment of State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, ChinaDepartment of Innovation Center for Diagnostics and Treatment of Thalassemia, Nanfang Hospital, Southern Medical University, Guangzhou, China; Department of Medical Genetics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, ChinaDepartment of Innovation Center for Diagnostics and Treatment of Thalassemia, Nanfang Hospital, Southern Medical University, Guangzhou, China; Department of Medical Genetics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, ChinaDepartment of Innovation Center for Diagnostics and Treatment of Thalassemia, Nanfang Hospital, Southern Medical University, Guangzhou, China; Department of Medical Genetics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, ChinaDepartment of Innovation Center for Diagnostics and Treatment of Thalassemia, Nanfang Hospital, Southern Medical University, Guangzhou, China; Department of Medical Genetics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, ChinaDepartment of Innovation Center for Diagnostics and Treatment of Thalassemia, Nanfang Hospital, Southern Medical University, Guangzhou, China; Department of Medical Genetics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, ChinaDepartment of Innovation Center for Diagnostics and Treatment of Thalassemia, Nanfang Hospital, Southern Medical University, Guangzhou, China; Department of Medical Genetics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, ChinaDepartment of State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, ChinaDepartment of Key Laboratory of Model Animals and Stem Cell Biology, Engineering Research Center of Reproduction and Translational Medicine, Guangxiu Hospital, Health Science Center, Hunan Normal University, Changsha, ChinaDepartment of Pediatrics, 923rd Hospital of the People's Liberation Army, Nanning, China; Xinhua Zhang, Department of Pediatrics, 923rd Hospital of the People's Liberation Army, No.52 Zhiwu Rd, Qingxiu District, Nanning, Guangxi Zhuang Autonomous Region, 530022, China;Department of Innovation Center for Diagnostics and Treatment of Thalassemia, Nanfang Hospital, Southern Medical University, Guangzhou, China; Department of Medical Genetics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China; Correspondence: Xiangmin Xu, Innovation Center for Diagnostics and Treatment of Thalassemia, Nanfang Hospital, Department of Medical Genetics, School of Basic Medical Sciences, Southern Medical University, 1023, South Shatai Rd, Baiyun District, Guangzhou, Guangdong, 510515, China;Abstract: β-Thalassemia is a common monogenic disorder caused by genetic defects in β-globin genes (HBB) resulting in imbalanced synthesis of α-/β-globin and ineffective erythropoiesis. It has been well documented that patients with β-thalassemia, or even carriers, mostly experience reactivation of fetal hemoglobin (Hb F), but its underlying mechanisms are incompletely understood. We took advantage of a previously established cohort of 1142 patients with β-thalassemia with diverse thalassemic mutations subjected to targeted next-generation sequencing. Genotype-phenotype association studies demonstrated that the HBB:c.-78A>G had a remarkable effect on the elevation of Hb F levels compared with other β-thalassemic mutations. To experimentally validate this conclusion, the ribonucleoprotein transfection complex through homology-directed repair by electroporation was performed, from which we observed a consistent increase of Hb F expression in both HUDEP-2 and primary CD34+ cell lines. Furthermore, chromatin immunoprecipitation–quantitative polymerase chain reaction, dual-luciferase reporter assay, and circular chromosome conformation capture (4C) assays validated a decreased occupancy of the HBB TATA box by TATA-binding protein (TBP), leading to boosted expression of γ-globin genes by enhanced interaction between locus control regions (LCRs) and γ-globin gene promoters. The patient-based investigation and experimental validations presented in this study might lead to a better understanding of stage-specific globin-gene expression mediated by competitive binding of distal enhancers (LCRs).http://www.sciencedirect.com/science/article/pii/S2473952925002095 |
| spellingShingle | Mengyang Song Xiaolei Wei Hualei Luo Jueheng Wang Yuhua Ye Lang Qin Chao Niu Yong Long Xingmin Wang Congwen Shao Miao Yu Feng Gu Xinhua Zhang Xiangmin Xu A common TBP-binding site mutation elevates fetal hemoglobin levels by competitive globin switching change in β-thalassemia Blood Advances |
| title | A common TBP-binding site mutation elevates fetal hemoglobin levels by competitive globin switching change in β-thalassemia |
| title_full | A common TBP-binding site mutation elevates fetal hemoglobin levels by competitive globin switching change in β-thalassemia |
| title_fullStr | A common TBP-binding site mutation elevates fetal hemoglobin levels by competitive globin switching change in β-thalassemia |
| title_full_unstemmed | A common TBP-binding site mutation elevates fetal hemoglobin levels by competitive globin switching change in β-thalassemia |
| title_short | A common TBP-binding site mutation elevates fetal hemoglobin levels by competitive globin switching change in β-thalassemia |
| title_sort | common tbp binding site mutation elevates fetal hemoglobin levels by competitive globin switching change in β thalassemia |
| url | http://www.sciencedirect.com/science/article/pii/S2473952925002095 |
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