title: Induction of adult levels of β-globin in human erythroid cells that intrinsically express embryonic or fetal globin by transduction with KLF1 and BCL11A-XL.
abstract: A major barrier to the clinical use of erythrocytes generated in vitro from pluripotent stem cells or cord blood progenitors is failure of these erythrocytes to express adult hemoglobin. The key regulators of globin switching KLF1 and BCL11A are absent or at a lower level than in adult cells in K562 and erythroid cells differentiated in vitro from induced pluripotent stem cells and cord blood progenitors. Transfection or transduction of K562 and cord blood erythroid cells with either KLF1 or BCL11A-XL had little effect on β-globin expression. In contrast, transduction with both transcription factors stimulated β-globin expression. Similarly, increasing the level of BCL11A-XL in the induced pluripotent stem cell-derived erythroid cell line HiDEP-1, which has levels of endogenous KLF1 similar to adult cells but lacks BCL11A, resulted in levels of β-globin equivalent to that of adult erythroid cells. Interestingly, this increase in β-globin was coincident with a decrease in ε- and ζ-, but not γ-globin, implicating BCL11A in repression of embryonic globin expression. The data show that KLF1 and BCL11A-XL together are required, but sufficient to induce adult levels of β-globin in induced pluripotent stem cell and cord blood-derived erythroid cells that intrinsically express embryonic or fetal globin.
authors.affiliation: School of Biochemistry, University of Bristol, Bristol, United Kingdom Department of Biochemistry, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
authors.forename: Kongtana
authors.initials: K
authors.lastname: Trakarnsanga
authors.affiliation: School of Biochemistry, University of Bristol, Bristol, United Kingdom.
authors.forename: Marieangela C
authors.initials: MC
authors.lastname: Wilson
authors.affiliation: School of Biochemistry, University of Bristol, Bristol, United Kingdom.
authors.forename: Winnie
authors.initials: W
authors.lastname: Lau
authors.affiliation: Bristol Institute for Transfusion Sciences, National Health Service Blood and Transplant (NHSBT), Filton, Bristol, UK.
authors.forename: Belinda K
authors.initials: BK
authors.lastname: Singleton
authors.affiliation: Bristol Institute for Transfusion Sciences, National Health Service Blood and Transplant (NHSBT), Filton, Bristol, UK.
authors.forename: Steve F
authors.initials: SF
authors.lastname: Parsons
authors.affiliation: School of Biochemistry, University of Bristol, Bristol, United Kingdom.
authors.forename: Punthita
authors.initials: P
authors.lastname: Sakuntanaga
authors.affiliation: Cell Engineering Division, RIKEN BioResource Center, Tsukuba, Ibaraki, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Kurita
authors.affiliation: Cell Engineering Division, RIKEN BioResource Center, Tsukuba, Ibaraki, Japan.
authors.forename: Yukio
authors.initials: Y
authors.lastname: Nakamura
authors.affiliation: Bristol Institute for Transfusion Sciences, National Health Service Blood and Transplant (NHSBT), Filton, Bristol, UK jan.frayne@bristol.ac.uk david.anstee@nhsbt.nhs.uk.
authors.forename: David J
authors.initials: DJ
authors.lastname: Anstee
authors.affiliation: School of Biochemistry, University of Bristol, Bristol, United Kingdom jan.frayne@bristol.ac.uk david.anstee@nhsbt.nhs.uk.
authors.forename: Jan
authors.initials: J
authors.lastname: Frayne
mesh_terms: D053687:Adult Stem Cells / Q000166:cytology / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics*; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D041905:Erythroid Cells / Q000166:cytology / Q000378:metabolism*; D005319:Fetal Hemoglobin / Q000235:genetics*; D015870:Gene Expression; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D020014:K562 Cells; D051741:Kruppel-Like Transcription Factors / Q000235:genetics*; D009687:Nuclear Proteins / Q000235:genetics*; D010641:Phenotype; D012097:Repressor Proteins; D014161:Transduction, Genetic; D014162:Transfection; D055544:beta-Globins / Q000235:genetics*; D055547:epsilon-Globins / Q000235:genetics; D055545:gamma-Globins / Q000235:genetics
chemical_list: C486563:BCL11A protein, human; D002352:Carrier Proteins; D051741:Kruppel-Like Transcription Factors; D009687:Nuclear Proteins; D012097:Repressor Proteins; D055544:beta-Globins; D055547:epsilon-Globins; C080774:erythroid Kruppel-like factor; D055545:gamma-Globins; D005319:Fetal Hemoglobin
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