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

Samuelson, Clare; Radtke, Stefan; Zhu, Haiying; Llewellyn, Mallory J; Fields, Emily; Cook, Savannah; Huang, Meei‐Li; Jerome, Keith R.; Kiem, Hans–Peter; Humbert, Olivier

doi:10.1016/j.omtm.2021.10.008

Cited by 28 publications

(27 citation statements)

References 48 publications

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“…In recent years, progress on the knowledge of molecular mechanisms involved in Hb switching, the concept of HPHF, combined with epidemiological and clinical observations has provided important evidence on the beneficial role of increasing HbF levels in ameliorating the clinical complications of β-thalassemia and SCD [17]. This can be achieved by either classic drug-modulating approaches or by gene therapy and genome-editing approaches [18][19][20][21][22][23][24]. Although encouraging and promising results have been reported on gene therapy/genome editing and HbF expression, their safety is still to be determined and their expected costs are to be considered as well [25][26][27].…”

Section: Introductionmentioning

confidence: 99%

Pharmacological Induction of Fetal Hemoglobin in β-Thalassemia and Sickle Cell Disease: An Updated Perspective

et al. 2022

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A significant amount of attention has recently been devoted to the mechanisms involved in hemoglobin (Hb) switching, as it has previously been established that the induction of fetal hemoglobin (HbF) production in significant amounts can reduce the severity of the clinical course in diseases such as β-thalassemia and sickle cell disease (SCD). While the induction of HbF using lentiviral and genome-editing strategies has been made possible, they present limitations. Meanwhile, progress in the use of pharmacologic agents for HbF induction and the identification of novel HbF-inducing strategies has been made possible as a result of a better understanding of γ-globin regulation. In this review, we will provide an update on all current pharmacological inducer agents of HbF in β-thalassemia and SCD in addition to the ongoing research into other novel, and potentially therapeutic, HbF-inducing agents.

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Section: Introductionmentioning

confidence: 99%

Pharmacological Induction of Fetal Hemoglobin in β-Thalassemia and Sickle Cell Disease: An Updated Perspective

et al. 2022

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“…Mutations of a single or several nucleotides in human genome are responsible for major hereditary health problems ( Benusiglio et al, 2021 ; Samuelson et al, 2021 ; Xiao et al, 2021 ; Chen et al, 2022 ; Cobo et al, 2022 ). To date, about 7,000 hereditary diseases are estimated to be caused by monogenic mutations ( Claussnitzer et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

Improvements of nuclease and nickase gene modification techniques for the treatment of genetic diseases

Mbakam²,

Song³

et al. 2022

Front. Genome Ed.

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Advancements in genome editing make possible to exploit the functions of enzymes for efficient DNA modifications with tremendous potential to treat human genetic diseases. Several nuclease genome editing strategies including Meganucleases (MNs), Zinc Finger Nucleases (ZFNs), Transcription Activator-like Effector Nucleases (TALENs) and Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR associated proteins (CRISPR-Cas) have been developed for the correction of genetic mutations. CRISPR-Cas has further been engineered to create nickase genome editing tools including Base editors and Prime editors with much precision and efficacy. In this review, we summarized recent improvements in nuclease and nickase genome editing approaches for the treatment of genetic diseases. We also highlighted some limitations for the translation of these approaches into clinical applications.

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“…To maximize HbF production, these CRISPR-Cas9-based gene editing approaches can be combined, as recently proposed by Han et al and by Samuelson et al [ 39 , 40 ], who reported on a multiplex gene editing strategy based on the combination of two single gene editing approaches, one aimed at silencing the BCL11A repressor, the other aimed at disrupting the BCL11A binding sites present within the γ-globin gene promoter. Another example of multiplex gene editing approaches is that recently published by Psatha et al [ 41 ], who studied the combination of CRISPR-Cas9-based cis and trans fetal globin reactivation mutations, demonstrating that this strategy leads to a significant increase in HbF production when comparison was performed with the single editing procedures.…”

Section: Introductionmentioning

confidence: 99%

“…The advantage of this strategy is that both protocols use the same target cells (CD34 + erythroid progenitors) and no differences are expected in the clinical steps to be followed for collecting the CD34 + cells to be gene edited and for preparing the patients for the infusion of gene-edited cells (e.g., stem cells collected via mobilization and apheresis, myeloablative conditioning, infusion of corrected stem cells for the engraftment and immune reconstitution) [ 29 ]. On the other hand, a major drawback is expected, i.e., higher off-targeting effects and genotoxicity [ 40 ]. Supporting a caution in using multiplexed CRISPR-Cas9-based approaches, Samuelson et al recently reported that multiplex CRISPR-Cas9 genome editing in hematopoietic stem cells for fetal hemoglobin reinduction generates chromosomal translocations [ 40 ].…”

Section: Introductionmentioning

confidence: 99%

“…On the other hand, a major drawback is expected, i.e., higher off-targeting effects and genotoxicity [ 40 ]. Supporting a caution in using multiplexed CRISPR-Cas9-based approaches, Samuelson et al recently reported that multiplex CRISPR-Cas9 genome editing in hematopoietic stem cells for fetal hemoglobin reinduction generates chromosomal translocations [ 40 ]. For these reasons, we therefore decided to use for HbF induction a repositioned drug, rapamycin [ 42 , 43 , 44 , 45 , 46 , 47 , 48 ], among those already validated and used in clinical trials [ 49 , 50 , 51 , 52 ].…”

Section: Introductionmentioning

confidence: 99%

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Co-Treatment of Erythroid Cells from β-Thalassemia Patients with CRISPR-Cas9-Based β039-Globin Gene Editing and Induction of Fetal Hemoglobin

Cosenza

Zuccato

Zurlo

et al. 2022

Genes

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Gene editing (GE) is an efficient strategy for correcting genetic mutations in monogenic hereditary diseases, including β-thalassemia. We have elsewhere reported that CRISPR-Cas9-based gene editing can be employed for the efficient correction of the β039-thalassemia mutation. On the other hand, robust evidence demonstrates that the increased production of fetal hemoglobin (HbF) can be beneficial for patients with β-thalassemia. The aim of our study was to verify whether the de novo production of adult hemoglobin (HbA) using CRISPR-Cas9 gene editing can be combined with HbF induction protocols. The gene editing of the β039-globin mutation was obtained using a CRISPR-Cas9-based experimental strategy; the correction of the gene sequence and the transcription of the corrected gene were analyzed by allele-specific droplet digital PCR and RT-qPCR, respectively; the relative content of HbA and HbF was studied by high-performance liquid chromatography (HPLC) and Western blotting. For HbF induction, the repurposed drug rapamycin was used. The data obtained conclusively demonstrate that the maximal production of HbA and HbF is obtained in GE-corrected, rapamycin-induced erythroid progenitors isolated from β039-thalassemia patients. In conclusion, GE and HbF induction might be used in combination in order to achieve the de novo production of HbA together with an increase in induced HbF.

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

Cited by 28 publications

References 48 publications

Pharmacological Induction of Fetal Hemoglobin in β-Thalassemia and Sickle Cell Disease: An Updated Perspective

Pharmacological Induction of Fetal Hemoglobin in β-Thalassemia and Sickle Cell Disease: An Updated Perspective

Improvements of nuclease and nickase gene modification techniques for the treatment of genetic diseases

Co-Treatment of Erythroid Cells from β-Thalassemia Patients with CRISPR-Cas9-Based β039-Globin Gene Editing and Induction of Fetal Hemoglobin

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