Sickle cell disease (SCD) is caused by a single amino acid change in the adult hemoglobin (Hb) beta chain that causes Hb polymerization and red blood cell (RBC) sickling. The co-inheritance of mutations causing fetal gamma-globin production in adult life hereditary persistence of fetal Hb (HPFH) reduces the clinical severity of SCD. HPFH mutations in the HBG gamma-globin promoters disrupt binding sites for the repressors BCL1 1A and LRF. We used CRISPR-Cas9 to mimic HPFH mutations in the HBG promoters by generating insertions and deletions, leading to disruption of known and putative repressor binding sites. Editing of the LRF-binding site in patient-derived hematopoietic stem/progenitor cells (HSPCs) resulted in gamma-globin derepression and correction of the sickling phenotype. Xenotransplantation of HSPCs treated with gRNAs targeting the LRF-binding site showed a high editing efficiency in repopulating HSPCs. This study identifies the LRF-binding site as a potent target for genome-editing treatment of SCD.

Editing a γ-globin repressor binding site restores fetal hemoglobin synthesis and corrects the sickle cell disease phenotype / Weber, Leslie; Frati, Giacomo; Felix, Tristan; Hardouin, Giulia; Casini, Antonio; Wollenschlaeger, Clara; Meneghini, Vasco; Masson, Cecile; De Cian, Anne; Chalumeau, Anne; Mavilio, Fulvio; Amendola, Mario; Andre-Schmutz, Isabelle; Cereseto, Anna; El Nemer, Wassim; Concordet, Jean-Paul; Giovannangeli, Carine; Cavazzana, Marina; Miccio, Annarita. - In: SCIENCE ADVANCES. - ISSN 2375-2548. - 6:7(2020). [10.1126/sciadv.aay9392]

Editing a γ-globin repressor binding site restores fetal hemoglobin synthesis and corrects the sickle cell disease phenotype

Meneghini, Vasco;
2020-01-01

Abstract

Sickle cell disease (SCD) is caused by a single amino acid change in the adult hemoglobin (Hb) beta chain that causes Hb polymerization and red blood cell (RBC) sickling. The co-inheritance of mutations causing fetal gamma-globin production in adult life hereditary persistence of fetal Hb (HPFH) reduces the clinical severity of SCD. HPFH mutations in the HBG gamma-globin promoters disrupt binding sites for the repressors BCL1 1A and LRF. We used CRISPR-Cas9 to mimic HPFH mutations in the HBG promoters by generating insertions and deletions, leading to disruption of known and putative repressor binding sites. Editing of the LRF-binding site in patient-derived hematopoietic stem/progenitor cells (HSPCs) resulted in gamma-globin derepression and correction of the sickling phenotype. Xenotransplantation of HSPCs treated with gRNAs targeting the LRF-binding site showed a high editing efficiency in repopulating HSPCs. This study identifies the LRF-binding site as a potent target for genome-editing treatment of SCD.
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.11768/135050
Citazioni
  • ???jsp.display-item.citation.pmc??? 44
  • Scopus 85
  • ???jsp.display-item.citation.isi??? 78
social impact