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Base-editing-mediated dissection of a γ-globin cis-regulatory element for the therapeutic reactivation of fetal hemoglobin expression

Author

Listed:
  • Panagiotis Antoniou

    (Université Paris Cité, Imagine Institute, Laboratory of chromatin and gene regulation during development, INSERM UMR 1163)

  • Giulia Hardouin

    (Université Paris Cité, Imagine Institute, Laboratory of chromatin and gene regulation during development, INSERM UMR 1163
    Université Paris Cité, Imagine Institute, Laboratory of Human Lymphohematopoiesis, INSERM UMR 1163
    Assistance Publique Hopitaux de Paris, INSERM)

  • Pierre Martinucci

    (Université Paris Cité, Imagine Institute, Laboratory of chromatin and gene regulation during development, INSERM UMR 1163)

  • Giacomo Frati

    (Université Paris Cité, Imagine Institute, Laboratory of chromatin and gene regulation during development, INSERM UMR 1163)

  • Tristan Felix

    (Université Paris Cité, Imagine Institute, Laboratory of chromatin and gene regulation during development, INSERM UMR 1163)

  • Anne Chalumeau

    (Université Paris Cité, Imagine Institute, Laboratory of chromatin and gene regulation during development, INSERM UMR 1163)

  • Letizia Fontana

    (Université Paris Cité, Imagine Institute, Laboratory of chromatin and gene regulation during development, INSERM UMR 1163)

  • Jeanne Martin

    (Université Paris Cité, Imagine Institute, Laboratory of chromatin and gene regulation during development, INSERM UMR 1163)

  • Cecile Masson

    (Bioinformatics Platform, Imagine Institute)

  • Megane Brusson

    (Université Paris Cité, Imagine Institute, Laboratory of chromatin and gene regulation during development, INSERM UMR 1163)

  • Giulia Maule

    (CIBIO, University of Trento)

  • Marion Rosello

    (Sorbonne Université, INSERM, CNRS, Institut de la Vision)

  • Carine Giovannangeli

    (INSERM U1154, CNRS UMR7196, Museum National d’Histoire Naturelle)

  • Vincent Abramowski

    (Université Paris Cité, Imagine Institute, Laboratory of genome dynamics in the immune system, INSERM UMR 1163)

  • Jean-Pierre de Villartay

    (Université Paris Cité, Imagine Institute, Laboratory of genome dynamics in the immune system, INSERM UMR 1163)

  • Jean-Paul Concordet

    (INSERM U1154, CNRS UMR7196, Museum National d’Histoire Naturelle)

  • Filippo Del Bene

    (Sorbonne Université, INSERM, CNRS, Institut de la Vision)

  • Wassim El Nemer

    (Établissement Français du Sang, UMR 7268
    Laboratoire d’Excellence GR-Ex)

  • Mario Amendola

    (Genethon
    Université Paris-Saclay, Univ Evry, Inserm, Genethon, Integrare research unit UMR_S951)

  • Marina Cavazzana

    (Assistance Publique Hopitaux de Paris, INSERM
    Université Paris Cité
    Imagine Institute)

  • Anna Cereseto

    (CIBIO, University of Trento)

  • Oriana Romano

    (University of Modena and Reggio Emilia)

  • Annarita Miccio

    (Université Paris Cité, Imagine Institute, Laboratory of chromatin and gene regulation during development, INSERM UMR 1163)

Abstract

Sickle cell disease and β-thalassemia affect the production of the adult β-hemoglobin chain. The clinical severity is lessened by mutations that cause fetal γ-globin expression in adult life (i.e., the hereditary persistence of fetal hemoglobin). Mutations clustering ~200 nucleotides upstream of the HBG transcriptional start sites either reduce binding of the LRF repressor or recruit the KLF1 activator. Here, we use base editing to generate a variety of mutations in the −200 region of the HBG promoters, including potent combinations of four to eight γ-globin-inducing mutations. Editing of patient hematopoietic stem/progenitor cells is safe, leads to fetal hemoglobin reactivation and rescues the pathological phenotype. Creation of a KLF1 activator binding site is the most potent strategy – even in long-term repopulating hematopoietic stem/progenitor cells. Compared with a Cas9-nuclease approach, base editing avoids the generation of insertions, deletions and large genomic rearrangements and results in higher γ-globin levels. Our results demonstrate that base editing of HBG promoters is a safe, universal strategy for treating β-hemoglobinopathies.

Suggested Citation

  • Panagiotis Antoniou & Giulia Hardouin & Pierre Martinucci & Giacomo Frati & Tristan Felix & Anne Chalumeau & Letizia Fontana & Jeanne Martin & Cecile Masson & Megane Brusson & Giulia Maule & Marion Ro, 2022. "Base-editing-mediated dissection of a γ-globin cis-regulatory element for the therapeutic reactivation of fetal hemoglobin expression," Nature Communications, Nature, vol. 13(1), pages 1-22, December.
  • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-34493-1
    DOI: 10.1038/s41467-022-34493-1
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    References listed on IDEAS

    as
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