Direct correction of haemoglobin E β-thalassaemia using base editors
Abstract Haemoglobin E (HbE) β-thalassaemia causes approximately 50% of all severe thalassaemia worldwide; equating to around 30,000 births per year. HbE β-thalassaemia is due to a point mutation in codon 26 of the human HBB gene on one allele (GAG; glutamatic acid → AAG; lysine, E26K), and any muta...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-04-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-023-37604-8 |
| _version_ | 1797840943142928384 |
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| author | Mohsin Badat Ayesha Ejaz Peng Hua Siobhan Rice Weijiao Zhang Lance D. Hentges Christopher A. Fisher Nicholas Denny Ron Schwessinger Nirmani Yasara Noemi B. A. Roy Fadi Issa Andi Roy Paul Telfer Jim Hughes Sachith Mettananda Douglas R. Higgs James O. J. Davies |
| author_facet | Mohsin Badat Ayesha Ejaz Peng Hua Siobhan Rice Weijiao Zhang Lance D. Hentges Christopher A. Fisher Nicholas Denny Ron Schwessinger Nirmani Yasara Noemi B. A. Roy Fadi Issa Andi Roy Paul Telfer Jim Hughes Sachith Mettananda Douglas R. Higgs James O. J. Davies |
| author_sort | Mohsin Badat |
| collection | DOAJ |
| description | Abstract Haemoglobin E (HbE) β-thalassaemia causes approximately 50% of all severe thalassaemia worldwide; equating to around 30,000 births per year. HbE β-thalassaemia is due to a point mutation in codon 26 of the human HBB gene on one allele (GAG; glutamatic acid → AAG; lysine, E26K), and any mutation causing severe β-thalassaemia on the other. When inherited together in compound heterozygosity these mutations can cause a severe thalassaemic phenotype. However, if only one allele is mutated individuals are carriers for the respective mutation and have an asymptomatic phenotype (β-thalassaemia trait). Here we describe a base editing strategy which corrects the HbE mutation either to wildtype (WT) or a normal variant haemoglobin (E26G) known as Hb Aubenas and thereby recreates the asymptomatic trait phenotype. We have achieved editing efficiencies in excess of 90% in primary human CD34 + cells. We demonstrate editing of long-term repopulating haematopoietic stem cells (LT-HSCs) using serial xenotransplantation in NSG mice. We have profiled the off-target effects using a combination of circularization for in vitro reporting of cleavage effects by sequencing (CIRCLE-seq) and deep targeted capture and have developed machine-learning based methods to predict functional effects of candidate off-target mutations. |
| first_indexed | 2024-04-09T16:22:59Z |
| format | Article |
| id | doaj.art-88dbd35ddfe1418a8b4cd19610b19c39 |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-04-09T16:22:59Z |
| publishDate | 2023-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-88dbd35ddfe1418a8b4cd19610b19c392023-04-23T11:23:25ZengNature PortfolioNature Communications2041-17232023-04-011411710.1038/s41467-023-37604-8Direct correction of haemoglobin E β-thalassaemia using base editorsMohsin Badat0Ayesha Ejaz1Peng Hua2Siobhan Rice3Weijiao Zhang4Lance D. Hentges5Christopher A. Fisher6Nicholas Denny7Ron Schwessinger8Nirmani Yasara9Noemi B. A. Roy10Fadi Issa11Andi Roy12Paul Telfer13Jim Hughes14Sachith Mettananda15Douglas R. Higgs16James O. J. Davies17MRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of OxfordMRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of OxfordMRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of OxfordMRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of OxfordMRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of OxfordMRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of OxfordMRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of OxfordMRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of OxfordMRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of OxfordDepartment of Paediatrics, University of KelaniyaMRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of OxfordTransplantation Research and Immunology Group, Nuffield Department of Surgical Sciences, University of OxfordMRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of OxfordDepartment of Clinical Haematology, Royal London Hospital, Barts Health NHS TrustMRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of OxfordDepartment of Paediatrics, University of KelaniyaLaboratory of Gene Regulation, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of OxfordMRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of OxfordAbstract Haemoglobin E (HbE) β-thalassaemia causes approximately 50% of all severe thalassaemia worldwide; equating to around 30,000 births per year. HbE β-thalassaemia is due to a point mutation in codon 26 of the human HBB gene on one allele (GAG; glutamatic acid → AAG; lysine, E26K), and any mutation causing severe β-thalassaemia on the other. When inherited together in compound heterozygosity these mutations can cause a severe thalassaemic phenotype. However, if only one allele is mutated individuals are carriers for the respective mutation and have an asymptomatic phenotype (β-thalassaemia trait). Here we describe a base editing strategy which corrects the HbE mutation either to wildtype (WT) or a normal variant haemoglobin (E26G) known as Hb Aubenas and thereby recreates the asymptomatic trait phenotype. We have achieved editing efficiencies in excess of 90% in primary human CD34 + cells. We demonstrate editing of long-term repopulating haematopoietic stem cells (LT-HSCs) using serial xenotransplantation in NSG mice. We have profiled the off-target effects using a combination of circularization for in vitro reporting of cleavage effects by sequencing (CIRCLE-seq) and deep targeted capture and have developed machine-learning based methods to predict functional effects of candidate off-target mutations.https://doi.org/10.1038/s41467-023-37604-8 |
| spellingShingle | Mohsin Badat Ayesha Ejaz Peng Hua Siobhan Rice Weijiao Zhang Lance D. Hentges Christopher A. Fisher Nicholas Denny Ron Schwessinger Nirmani Yasara Noemi B. A. Roy Fadi Issa Andi Roy Paul Telfer Jim Hughes Sachith Mettananda Douglas R. Higgs James O. J. Davies Direct correction of haemoglobin E β-thalassaemia using base editors Nature Communications |
| title | Direct correction of haemoglobin E β-thalassaemia using base editors |
| title_full | Direct correction of haemoglobin E β-thalassaemia using base editors |
| title_fullStr | Direct correction of haemoglobin E β-thalassaemia using base editors |
| title_full_unstemmed | Direct correction of haemoglobin E β-thalassaemia using base editors |
| title_short | Direct correction of haemoglobin E β-thalassaemia using base editors |
| title_sort | direct correction of haemoglobin e β thalassaemia using base editors |
| url | https://doi.org/10.1038/s41467-023-37604-8 |
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