TALEN-Mediated Gene Editing of HBG in Human Hematopoietic Stem Cells Leads to Therapeutic Fetal Hemoglobin Induction
In: Molecular Therapy: Methods & Clinical Development, Jg. 12 (2019-03-01), Heft 175-183, S. 175-183
Online
academicJournal
Zugriff:
Elements within the γ-hemoglobin promoters (HBG1 and HBG2) function to bind transcription complexes that mediate repression of fetal hemoglobin expression. Sickle cell disease (SCD) subjects with a 13-bp deletion in the HBG1 promoter exhibit a clinically favorable hereditary persistence of fetal hemoglobin (HPFH) phenotype. We developed TALENs targeting the homologous HBG promoters to de-repress fetal hemoglobin. Transfection of human CD34+ cells with TALEN mRNA resulted in indel generation in HBG1 (43%) and HBG2 (74%) including the 13-bp HPFH deletion (∼6%). Erythroid differentiation of edited cells revealed a 4.6-fold increase in γ-hemoglobin expression as detected by HPLC. Assessment of TALEN-edited CD34+ cells in vivo in a humanized mouse model demonstrated sustained presence of indels in hematopoietic cells up to 24 weeks. Indel rates remained unchanged following secondary transplantation consistent with editing of long-term repopulating stem cells (LT-HSCs). Human γ-hemoglobin expressing F cells were detected by flow cytometry approximately 50% more frequently in edited animals compared to mock. Together, these findings demonstrate that TALEN-mediated indel generation in the γ-hemoglobin promoter leads to high levels of fetal hemoglobin expression in vitro and in vivo, suggesting that this approach can provide therapeutic benefit in patients with SCD or β-thalassemia. Keywords: TALEN, hemoglobinopathy, HBG, hemoglobin, HPFH, SCD, sickle cell disease, thalassemia, gene editing, HSC
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TALEN-Mediated Gene Editing of HBG in Human Hematopoietic Stem Cells Leads to Therapeutic Fetal Hemoglobin Induction
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Autor/in / Beteiligte Person: | Lux, Christopher T. ; Pattabhi, Sowmya ; Berger, Mason ; Nourigat, Cynthia ; Flowers, David A. ; Negre, Olivier ; Humbert, Olivier ; Yang, Julia G. ; Lee, Calvin ; Jacoby, Kyle ; Bernstein, Irwin ; Kiem, Hans-Peter ; Scharenberg, Andrew ; Rawlings, David J. |
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Zeitschrift: | Molecular Therapy: Methods & Clinical Development, Jg. 12 (2019-03-01), Heft 175-183, S. 175-183 |
Veröffentlichung: | Elsevier, 2019 |
Medientyp: | academicJournal |
ISSN: | 2329-0501 (print) |
DOI: | 10.1016/j.omtm.2018.12.008 |
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