Dual alpha-globin-truncated erythropoietin receptor knockin restores hemoglobin production in alpha-thalassemia-derived erythroid cells.
Publication Year:
2025
PubMed ID:
39754719
Funding Grants:
Public Summary:
The most severe form of α-thalassemia results from loss of all four copies of α-globin. Postnatally, patients face challenges similar to β-thalassemia, including severe anemia and erythrotoxicity due to the imbalance of β-globin and α-globin chains. Despite progress in genome editing treatments for β-thalassemia, there is no analogous curative option for α-thalassemia. To address this, we designed a Cas9/AAV6-mediated genome editing strategy that integrates a functional α-globin gene into the β-globin locus in α-thalassemia patient-derived hematopoietic stem and progenitor cells (HSPCs). Incorporation of a truncated erythropoietin receptor transgene into the α-globin integration cassette significantly increased erythropoietic output from edited HSPCs and led to the most robust production of α-globin, and consequently hemoglobin tetramers. By directing edited HSPCs toward increased production of clinically relevant erythroid cells, this approach has the potential to mitigate the limitations of current treatments for the hemoglobinopathies, including low genome editing and low engraftment rates.
Scientific Abstract:
The most severe form of alpha-thalassemia results from loss of all four copies of alpha-globin. Postnatally, patients face challenges similar to beta-thalassemia, including severe anemia and erythrotoxicity due to the imbalance of beta-globin and alpha-globin chains. Despite progress in genome editing treatments for beta-thalassemia, there is no analogous curative option for alpha-thalassemia. To address this, we designed a Cas9/AAV6-mediated genome editing strategy that integrates a functional alpha-globin gene into the beta-globin locus in alpha-thalassemia patient-derived hematopoietic stem and progenitor cells (HSPCs). Incorporation of a truncated erythropoietin receptor transgene into the alpha-globin integration cassette significantly increased erythropoietic output from edited HSPCs and led to the most robust production of alpha-globin, and consequently hemoglobin tetramers. By directing edited HSPCs toward increased production of clinically relevant erythroid cells, this approach has the potential to mitigate the limitations of current treatments for the hemoglobinopathies, including low genome editing and low engraftment rates.
