Multiplex CRISPR/Cas9 genome editing in hematopoietic stem cells for fetal hemoglobin reinduction generates chromosomal translocations

Sickle cell disease and β-thalassemia are common monogenic disorders that cause significant morbidity and mortality globally. The only curative treatment currently is allogeneic hematopoietic stem cell transplantation, which is unavailable to many patients due to a lack of matched donors and carries...

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Autores principales:	Clare Samuelson, Stefan Radtke, Haiying Zhu, Mallory Llewellyn, Emily Fields, Savannah Cook, Meei-Li W. Huang, Keith R. Jerome, Hans-Peter Kiem, Olivier Humbert
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	BCL11A CRISPR/Cas9 fetal hemoglobin genome editing HBG multiplex editing Genetics QH426-470 Cytology QH573-671
Acceso en línea:	https://doaj.org/article/c920282311d94065aac1d4319ebc7ffb
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spelling	oai:doaj.org-article:c920282311d94065aac1d4319ebc7ffb2021-11-20T05:06:43ZMultiplex CRISPR/Cas9 genome editing in hematopoietic stem cells for fetal hemoglobin reinduction generates chromosomal translocations2329-050110.1016/j.omtm.2021.10.008https://doaj.org/article/c920282311d94065aac1d4319ebc7ffb2021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2329050121001650https://doaj.org/toc/2329-0501Sickle cell disease and β-thalassemia are common monogenic disorders that cause significant morbidity and mortality globally. The only curative treatment currently is allogeneic hematopoietic stem cell transplantation, which is unavailable to many patients due to a lack of matched donors and carries risks including graft-versus-host disease. Genome editing therapies targeting either the BCL11A erythroid enhancer or the HBG promoter are already demonstrating success in reinducing fetal hemoglobin. However, where a single locus is targeted, reliably achieving levels high enough to deliver an effective cure remains a challenge. We investigated the application of a CRISPR/Cas9 multiplex genome editing approach, in which both the BCL11A erythroid enhancer and HBG promoter are disrupted within human hematopoietic stem cells. We demonstrate superior fetal hemoglobin reinduction with this dual-editing approach without compromising engraftment or lineage differentiation potential of edited cells post-xenotransplantation. However, multiplex editing consistently resulted in the generation of chromosomal rearrangement events that persisted in vivo following transplantation into immunodeficient mice. The risk of oncogenic events resulting from such translocations therefore currently prohibits its clinical translation, but it is anticipated that, in the future, alternative editing platforms will help alleviate this risk.Clare SamuelsonStefan RadtkeHaiying ZhuMallory LlewellynEmily FieldsSavannah CookMeei-Li W. HuangKeith R. JeromeHans-Peter KiemOlivier HumbertElsevierarticleBCL11ACRISPR/Cas9fetal hemoglobingenome editingHBGmultiplex editingGeneticsQH426-470CytologyQH573-671ENMolecular Therapy: Methods & Clinical Development, Vol 23, Iss , Pp 507-523 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	BCL11A CRISPR/Cas9 fetal hemoglobin genome editing HBG multiplex editing Genetics QH426-470 Cytology QH573-671
spellingShingle	BCL11A CRISPR/Cas9 fetal hemoglobin genome editing HBG multiplex editing Genetics QH426-470 Cytology QH573-671 Clare Samuelson Stefan Radtke Haiying Zhu Mallory Llewellyn Emily Fields Savannah Cook Meei-Li W. Huang Keith R. Jerome Hans-Peter Kiem Olivier Humbert Multiplex CRISPR/Cas9 genome editing in hematopoietic stem cells for fetal hemoglobin reinduction generates chromosomal translocations
description	Sickle cell disease and β-thalassemia are common monogenic disorders that cause significant morbidity and mortality globally. The only curative treatment currently is allogeneic hematopoietic stem cell transplantation, which is unavailable to many patients due to a lack of matched donors and carries risks including graft-versus-host disease. Genome editing therapies targeting either the BCL11A erythroid enhancer or the HBG promoter are already demonstrating success in reinducing fetal hemoglobin. However, where a single locus is targeted, reliably achieving levels high enough to deliver an effective cure remains a challenge. We investigated the application of a CRISPR/Cas9 multiplex genome editing approach, in which both the BCL11A erythroid enhancer and HBG promoter are disrupted within human hematopoietic stem cells. We demonstrate superior fetal hemoglobin reinduction with this dual-editing approach without compromising engraftment or lineage differentiation potential of edited cells post-xenotransplantation. However, multiplex editing consistently resulted in the generation of chromosomal rearrangement events that persisted in vivo following transplantation into immunodeficient mice. The risk of oncogenic events resulting from such translocations therefore currently prohibits its clinical translation, but it is anticipated that, in the future, alternative editing platforms will help alleviate this risk.
format	article
author	Clare Samuelson Stefan Radtke Haiying Zhu Mallory Llewellyn Emily Fields Savannah Cook Meei-Li W. Huang Keith R. Jerome Hans-Peter Kiem Olivier Humbert
author_facet	Clare Samuelson Stefan Radtke Haiying Zhu Mallory Llewellyn Emily Fields Savannah Cook Meei-Li W. Huang Keith R. Jerome Hans-Peter Kiem Olivier Humbert
author_sort	Clare Samuelson
title	Multiplex CRISPR/Cas9 genome editing in hematopoietic stem cells for fetal hemoglobin reinduction generates chromosomal translocations
title_short	Multiplex CRISPR/Cas9 genome editing in hematopoietic stem cells for fetal hemoglobin reinduction generates chromosomal translocations
title_full	Multiplex CRISPR/Cas9 genome editing in hematopoietic stem cells for fetal hemoglobin reinduction generates chromosomal translocations
title_fullStr	Multiplex CRISPR/Cas9 genome editing in hematopoietic stem cells for fetal hemoglobin reinduction generates chromosomal translocations
title_full_unstemmed	Multiplex CRISPR/Cas9 genome editing in hematopoietic stem cells for fetal hemoglobin reinduction generates chromosomal translocations
title_sort	multiplex crispr/cas9 genome editing in hematopoietic stem cells for fetal hemoglobin reinduction generates chromosomal translocations
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/c920282311d94065aac1d4319ebc7ffb
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Multiplex CRISPR/Cas9 genome editing in hematopoietic stem cells for fetal hemoglobin reinduction generates chromosomal translocations

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