A prime editing strategy to rewrite the γ-globin promoters and reactivate fetal hemoglobin for sickle cell disease
Abstract
Abstract Fetal hemoglobin reactivation is a promising therapy for β-hemoglobinopathies. We developed a prime editing strategy that introduces multiple mutations in the fetal γ-globin promoters that are expected to increase their activity. We tested multiple targets and optimized a variety of parameters to achieve ∼50% of precise edits in a hematopoietic cell line, with minimal off-target effects. This work improved our understanding of the complex DNA repair mechanisms involved in prime editing. We tested this strategy in patients’ hematopoietic stem/progenitor cells. Although editing efficiency was variable among donors, erythroid clones carrying multiple mutations expressed a significantly higher γ-globin level than cells carrying individual mutations, confirming the potential therapeutic benefit of our combined strategy for patients with β-hemoglobinopathies.
Article Details
Authors (14)
Anne Chalumeau
Maria Bou Dames
1Université Paris Cité, Imagine Institute, Laboratory of Chromatin and Gene Regulation during Development, INSERM Unité Mixte de Recherche 1163, Paris, France
Letizia Fontana
1Université Paris Cité, Imagine Institute, Laboratory of Chromatin and Gene Regulation during Development, INSERM Unité Mixte de Recherche 1163, Paris, France
Simone Amistadi
1Université Paris Cité, Imagine Institute, Laboratory of Chromatin and Gene Regulation during Development, INSERM Unité Mixte de Recherche 1163, Paris, France
Panagiotis Antoniou
Priyanka Loganathan
1Université Paris Cité, Imagine Institute, Laboratory of Chromatin and Gene Regulation during Development, INSERM Unité Mixte de Recherche 1163, Paris, France
Margaux Mombled
3Genethon, INSERM Unité Mixte de Recherche S951, Evry, France
Guillaume Corre
3Genethon, INSERM Unité Mixte de Recherche S951, Evry, France
Martin Peterka
Mario Amendola
3Genethon, INSERM Unité Mixte de Recherche S951, Evry, France
Carine Giovannangeli
Marcello Maresca
Annarita Miccio
Mégane Brusson