Combination of lentiviral and genome editing technologies for the treatment of sickle cell disease

Ramadier, Sophie; Chalumeau, Anne; Félix, Tristan; Othman, Nadia; Aknoun, Shérazade; Casini, Antonio; Maule, Giulia; Masson, Cécile; Cian, Anne De; Frati, Giacomo; Brusson, Mégane; Concordet, Jean-Paul; Cavazzana, Marina; Cereseto, Anna; Nemer, Wassim El; Amendola, Mario; Wattellier, Benoît; Meneghini, Vasco; Miccio, Annarita

doi:10.1016/j.ymthe.2021.08.019

Cited by 8 publications

(7 citation statements)

References 75 publications

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“…In both cases, the protocols were based on base-editing knock down the sickle β-globin in association with de novo expression of anti-sickling globin (AS3) or anti-sickling fetal γ-globins. While in this study expression of anti-sickling fetal γ-globins was obtained by gene addition ( Ramadier et al, 2022 ), this can be also obtained by gene editing procedures for induction of γ-globin gene expression, as already described in Figure 2 and recently reported in the study by Venkatesan et al 2023b) . These studies confirmed also the possible use of combined protcols based on complementary gene editing procedures.…”

Section: Combined Protocols Based On Gene Therapy and Gene Editingmentioning

confidence: 84%

“…In this respect, the combination of gene therapy and gene editing was studied by Ramadier et al, who described two therapeutic approaches combining LV-based gene addition therapy ansd CRISPR-Cas9 gene editing ( Ramadier et al, 2022 ). In both cases, the protocols were based on base-editing knock down the sickle β-globin in association with de novo expression of anti-sickling globin (AS3) or anti-sickling fetal γ-globins.…”

Section: Combined Protocols Based On Gene Therapy and Gene Editingmentioning

confidence: 99%

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Combined approaches for increasing fetal hemoglobin (HbF) and de novo production of adult hemoglobin (HbA) in erythroid cells from β-thalassemia patients: treatment with HbF inducers and CRISPR-Cas9 based genome editing

Finotti

Gambari

2023

Front. Genome Ed.

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Genome editing (GE) is one of the most efficient and useful molecular approaches to correct the effects of gene mutations in hereditary monogenetic diseases, including β-thalassemia. CRISPR-Cas9 gene editing has been proposed for effective correction of the β-thalassemia mutation, obtaining high-level “de novo” production of adult hemoglobin (HbA). In addition to the correction of the primary gene mutations causing β-thalassemia, several reports demonstrate that gene editing can be employed to increase fetal hemoglobin (HbF), obtaining important clinical benefits in treated β-thalassemia patients. This important objective can be achieved through CRISPR-Cas9 disruption of genes encoding transcriptional repressors of γ-globin gene expression (such as BCL11A, SOX6, KLF-1) or their binding sites in the HBG promoter, mimicking non-deletional and deletional HPFH mutations. These two approaches (β-globin gene correction and genome editing of the genes encoding repressors of γ-globin gene transcription) can be, at least in theory, combined. However, since multiplex CRISPR-Cas9 gene editing is associated with documented evidence concerning possible genotoxicity, this review is focused on the possibility to combine pharmacologically-mediated HbF induction protocols with the “de novo” production of HbA using CRISPR-Cas9 gene editing.

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Section: Combined Protocols Based On Gene Therapy and Gene Editingmentioning

confidence: 84%

Section: Combined Protocols Based On Gene Therapy and Gene Editingmentioning

confidence: 99%

Combined approaches for increasing fetal hemoglobin (HbF) and de novo production of adult hemoglobin (HbA) in erythroid cells from β-thalassemia patients: treatment with HbF inducers and CRISPR-Cas9 based genome editing

Finotti

Gambari

2023

Front. Genome Ed.

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“…VCN/cell is indicated below the graph. HD samples (HD donors) are HD HSPCs that were either mock-transfected (HBB WT) or modified using a CRISPR/Cas9-gRNA RNP complex disrupting the HBB gene (HBB KO) (Ramadier et al, 2021). Cells were differentiated toward the erythroid lineage.…”

Section: Resultsmentioning

confidence: 99%

“…Genome editing strategies have also been developed by several groups including ours (Antoniani et al, 2018; Ramadier et al, 2021; Weber et al, 2020) with early, promising clinical results (Frangoul et al, 2020) but long-term efficacy and safety needs to be demonstrated, particularly in light of the recent findings on the CRISPR/Cas9-induced genotoxicity (Boutin et al, 2022; Nahmad et al, 2022). On the contrary, our approach can be readily implemented in clinically accepted LV design and eventually translated to the bedside after preclinical biosafety and biodistribution studies, to change the goal of current gene therapy clinical trial from an amelioration of clinical signs and reduction of blood transfusion to a complete cure of the disease.…”

Section: Discussionmentioning

confidence: 99%

“…Quantitative phase images of label-free RBCs were taken using the SID4 HR GE camera (Phasics, Saint-Aubin, France) with an inverted microscope (ECLISPE Ti-E, Nikon) and a x40/0.60 objective. For each image, a segmentation procedure was performed using the BIO-Data R&D software (version 2.7.1.46) to isolate individual RBCs (Ramadier et al, 2021). For β-thalassemia samples, enucleated RBCs were selected based on their surface density (dry mass/surface), discarding nucleated cells with a surface density (pg/mm 2 ) >0.376 (βAS3) and >0.4 (βAS3/miRBCL11A) for β + /β + #1 and >0.358 (Mock), >0.368 (βAS3), and >0.365 (βAS3/miRBCL11A) for β 0 /β 0 #2 .…”

Section: Methodsmentioning

confidence: 99%

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Novel lentiviral vectors for gene therapy of sickle cell disease combining gene addition and gene silencing strategies

Brusson

Chalumeau

Martinucci

et al. 2022

Preprint

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Sickle cell disease (SCD) is due to a mutation in the β-globin (HBB) gene causing the production of the toxic sickle hemoglobin (HbS, α2βS2). Transplantation of autologous hematopoietic stem/progenitor cells (HSPCs) transduced with lentiviral vectors (LVs) expressing an anti-sickling β-globin (βAS) is a promising treatment; however, it is only partially effective and patients still present elevated HbS levels. Here, we developed a bifunctional LV expressing βAS3-globin and an artificial microRNA (amiR) specifically downregulating βS-globin expression with the aim of reducing HbS levels and favoring βAS3 incorporation into Hb tetramers. Efficient transduction of SCD HSPC by the bifunctional LV led to a substantial decrease of βS-globin transcripts in HSPC-derived erythroid cells, a significant reduction of HbS+ red cells and effective correction of the sickling phenotype, outperforming βAS gene addition and BCL11A gene silencing strategies. The bifunctional LV showed a standard integration profile and neither the HSPC viability, engraftment and multi-lineage differentiation nor the erythroid transcriptome and miRNAome were affected by the treatment, confirming the safety of this therapeutic strategy. In conclusion, the combination of gene addition and gene silencing strategies can improve the efficacy of current LV-based therapeutic approaches without increasing the mutagenic vector load, thus representing a novel treatment for SCD.

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CRISPR technology in human diseases

Feng,

Li,

Zhou

et al. 2024

MedComm

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Gene editing is a growing gene engineering technique that allows accurate editing of a broad spectrum of gene‐regulated diseases to achieve curative treatment and also has the potential to be used as an adjunct to the conventional treatment of diseases. Gene editing technology, mainly based on clustered regularly interspaced palindromic repeats (CRISPR)–CRISPR‐associated protein systems, which is capable of generating genetic modifications in somatic cells, provides a promising new strategy for gene therapy for a wide range of human diseases. Currently, gene editing technology shows great application prospects in a variety of human diseases, not only in therapeutic potential but also in the construction of animal models of human diseases. This paper describes the application of gene editing technology in hematological diseases, solid tumors, immune disorders, ophthalmological diseases, and metabolic diseases; focuses on the therapeutic strategies of gene editing technology in sickle cell disease; provides an overview of the role of gene editing technology in the construction of animal models of human diseases; and discusses the limitations of gene editing technology in the treatment of diseases, which is intended to provide an important reference for the applications of gene editing technology in the human disease.

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Combination of lentiviral and genome editing technologies for the treatment of sickle cell disease

Cited by 8 publications

References 75 publications

Combined approaches for increasing fetal hemoglobin (HbF) and de novo production of adult hemoglobin (HbA) in erythroid cells from β-thalassemia patients: treatment with HbF inducers and CRISPR-Cas9 based genome editing

Combined approaches for increasing fetal hemoglobin (HbF) and de novo production of adult hemoglobin (HbA) in erythroid cells from β-thalassemia patients: treatment with HbF inducers and CRISPR-Cas9 based genome editing

Novel lentiviral vectors for gene therapy of sickle cell disease combining gene addition and gene silencing strategies

CRISPR technology in human diseases

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