Disruption of the MBD2-NuRD complex but not MBD3-NuRD induces high level HbF expression in human adult erythroid cells
| Autor: | Gordon D. Ginder, Xia Li, Yukio Nakamura, David C. Williams, Alexander Azzo, Xiaofei Yu, Mikhail Dozmorov, Stephanie M. Bilinovich, Ryo Kurita |
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| Rok vydání: | 2019 |
| Předmět: |
Adult
ATP Binding Cassette Transporter Subfamily B 03 medical and health sciences 0302 clinical medicine Erythroid Cells Fetal hemoglobin Gene expression Humans Gene silencing gamma-Globins Cells Cultured Fetal Hemoglobin Gene knockdown Chemistry Hematology Chromatin Assembly and Disassembly Mi-2/NuRD complex 3. Good health Chromatin Cell biology DNA-Binding Proteins Editorial Mutation Histone deacetylase CHD4 Mi-2 Nucleosome Remodeling and Deacetylase Complex 030215 immunology |
| Zdroj: | Haematologica. 104:2361-2371 |
| ISSN: | 1592-8721 0390-6078 |
| DOI: | 10.3324/haematol.2018.210963 |
| Popis: | As high fetal hemoglobin levels ameliorate the underlying pathophysiological defects in sickle cell anemia and beta (β)-thalassemia, understanding the mechanisms that enforce silencing of fetal hemoglobin postnatally offers the promise of effective molecular therapy. Depletion of the Nucleosome Remodeling and Deacetylase complex member MBD2 causes a 10-20-fold increase in γ-globin gene expression in adult β-globin locus yeast artificial chromosome transgenic mice. To determine the effect of MBD2 depletion in human erythroid cells, genome editing technology was utilized to knockout MBD2 in Human Umbilical cord Derived Erythroid Progenitor-2 cells resulting in γ/γ+β mRNA levels of approximately 50% and approximately 40% fetal hemoglobin by high performance liquid chromatography. In contrast, MBD3 knockout had no appreciable effect on γ-globin expression. Knockdown of MBD2 in primary adult erythroid cells consistently increased γ/γ+β mRNA ratios by approximately 10-fold resulting in approximately 30-40% γ/γ+β mRNA levels and a corresponding increase in γ-globin protein. MBD2 exerts its repressive effects through recruitment of the chromatin remodeler CHD4 via a coiled-coil domain, and the histone deacetylase core complex via an intrinsically disordered region. Enforced expression of wild-type MBD2 in MBD2 knockout cells caused a 5-fold decrease in γ-globin mRNA while neither the coiled-coil mutant nor the intrinsically disordered region mutant MBD2 proteins had an inhibitory effect. Co-immunoprecipitation assays showed that the coiled-coil and intrinsically disorder region mutations disrupt complex formation by dissociating the CHD4 and the histone deacetylase core complex components, respectively. These results establish the MBD2 Nucleosome Remodeling and Deacetylase complex as a major silencer of fetal hemoglobin in human erythroid cells and point to the coiled-coil and intrinsically disordered region of MBD2 as potential therapeutic targets. |
| Databáze: | OpenAIRE |
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