Inclusion of a short hairpin RNA targeting &lt;i&gt;BCL11A&lt;/i&gt; into a β-globin expressing vector allows concurrent synthesis of curative adult and fetal hemoglobin

Lourenço, Sílvia Pires; Jarocha, Danuta; Ghiaccio, Valentina; Guerra, Amaliris; Abdulmalik, Osheiza; La, Ping; Zezulin, Alexandra U.; Smith‐Whitley, Kim; Kwiatkowski, Janet L.; Guzikowski, Virginia; Nakamura, Yukio; Raabe, Tobias; Breda, Laura; Rivella, Stefano

doi:10.3324/haematol.2020.276634

Cited by 6 publications

(9 citation statements)

References 15 publications

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“…At similar VCN, γ-globin levels were modest compared to those obtained by only expressing the shmiR targeting BCL11A-XL under the control of similar regulatory elements (Brendel et al, 2020; Esrick et al, 2021), suggesting that the processing of the intron-embedded amiR is less efficient or that βAS3 globin competes with γ-globin for the incorporation in the Hb tetramers – at least in SCD cells. Interestingly, our results are in line with a recent study describing a similar LV expressing simultaneously the T87Q β-globin (carrying only one anti-sickling amino acid) and an amiR targeting BCL11A-XL (Pires Lourenco et al, 2021). Of note, in the case of β-thalassemia, this strategy could be combined with the introduction of a second amiR targeting the α-globin, which was recently shown to improve the α/β ratio in erythrocytes derived from β-thalassemia HSPCs (Nualkaew et al, 2021).…”

Section: Discussionsupporting

confidence: 92%

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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Section: Discussionsupporting

confidence: 92%

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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“…Combined gene insertion and editing strategies are also emerging and would hopefully show synergistic effects that could better guarantee the desired hematologic response. For example, a new approach is currently being evaluated based on the inclusion of a short hairpin RNA targeting BCL11A into a β-globin expressing vector to allow concurrent synthesis of curative adult and fetal Hb 105. Despite the large array of pharmacologic agents in development,the goals of therapy remain the same: transfusion reduction in TDT and improvement of Hb level in NTDT.…”

mentioning

confidence: 99%

2021 update on clinical trials in β‐thalassemia

Musallam¹,

Bou‐Fakhredin

Cappellini

et al. 2021

American J Hematol

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The treatment landscape for patients with β-thalassemia is witnessing a swift evolution, yet several unmet needs continue to persist. Patients with transfusiondependent β-thalassemia (TDT) primarily rely on regular transfusion and iron chelation therapy, which can be associated with considerable treatment burden and cost.Patients with non-transfusion-dependent β-thalassemia (NTDT) are also at risk of significant morbidity due to the underlying anemia and iron overload, but treatment options in this patient subgroup are limited. In this review, we provide updates on clinical trials of novel therapies targeting the underlying pathology in β-thalassemia, including the α/non-α-globin chain imbalance, ineffective erythropoiesis, and iron dysregulation. | INTRODUCTIONThe β-thalassemias are a group of inherited disorders of hemoglobin (Hb) synthesis characterized by chronic anemia of varying severity.The degree of anemia relies on several genetic and environmental factors and determines the need for regular transfusion therapy. It is now common practice to classify patients as having transfusion-The TDT patients (β-thalassemia major and severe forms of HbE/β-thalassemia) are those who commonly present in early childhood with severe anemia and require lifelong transfusion therapy for survival. 1 Although the introduction of transfusions improved survival in TDT patients, it did not come without its own side-effect, systemic iron overload leading to end-organ damage and increased mortality from cardiac or hepatic disease. [2][3][4] Advances in iron chelation therapy and the introduction of MRI techniques to detect organ-specific iron overload have led to improved management and patient outcomes. 5,6 Still, TDT comes with considerable burden to the patient, clinician, and overall healthcare system owing to persistent morbidity and high healthcare utilization, poor access to optimal care and high treatment cost especially in resource-limited countries, and several unmet needs in terms of efficacy, safety and adherence to conventional therapies. 7 Allogeneic hematopoietic stem-cell transplantation (HSCT) has been used successfully for the past few decades to offer curative therapy for patients with TDT, but is only available to a minority of patients with compatible donors. 1 Patients with NTDT (β-thalassemia intermedia and mildmoderate forms of HbE/β-thalassemia) usually present later in childhood or even in adolescence with mild-moderate anemia that does not require immediate placement on a regular transfusion program. 1,8 Progress made over the past few decades has indicated that the diagnosis of NTDT carries greater morbidity than previously recognized.Ineffective erythropoiesis and anemia have been linked to an array of morbidities stemming from chronic hypoxia and an established hypercoagulable state. 1 Patients with Hb levels < 10 g/dl are at an increased risk of morbidity development, and variations of 1 g/dl can change a patient's morbidity risk. 9,10 There are currently no approved agents for the management of anem...

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“… 80 , 148 Alternatively, the transduction of HSC with lentiviral (LV) vectors harboring short hairpin RNA (shRNAs) has emerged as an effective delivery strategy for the clinical translation of RNAi‐based therapies. 149 , 150 , 151 , 152 In this strategy, the artificial miRNA scaffold is used to express a shRNA in the cell type of interest. 151 , 152 , 153 , 154 , 155 Such a design principle permits the shRNA to be processed in the same pathway as natural miRNAs and mediate silencing of target mRNA.…”

Section: Novel Therapies Targeting α‐Globin Expressionmentioning

confidence: 99%

“… 149 , 150 , 151 , 152 In this strategy, the artificial miRNA scaffold is used to express a shRNA in the cell type of interest. 151 , 152 , 153 , 154 , 155 Such a design principle permits the shRNA to be processed in the same pathway as natural miRNAs and mediate silencing of target mRNA. These findings signify important advances in lineage‐specific gene silencing, as this approach can be used as a flexible tool for the analysis of gene function and the development of gene‐specific therapeutics.…”

Section: Novel Therapies Targeting α‐Globin Expressionmentioning

confidence: 99%

“…Recent studies have demonstrated the beneficiary effects of intronic shRNA expression systems with coordinated β‐globin expression. 151 , 152 In this context, the generation of a lentiviral gene therapy vector comprised of the therapeutic anti‐sickling β A‐T87Q ‐globin gene was configured with an intronic miR30‐shRNA tailored to specifically reduce α2‐globin mRNA expression and correct the α/β‐globin chain imbalance in β‐thalassemia. 151 This approach has the important advantage of not only expressing the therapeutic β‐globin gene but also reducing excess α‐globin expression.…”

Section: Novel Therapies Targeting α‐Globin Expressionmentioning

confidence: 99%

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Interplay between α‐thalassemia and β‐hemoglobinopathies: Translating genotype–phenotype relationships into therapies

Vadolas,

Nualkaew,

Voon

et al. 2024

HemaSphere

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Abstractα‐Thalassemia represents one of the most important genetic modulators of β‐hemoglobinopathies. During this last decade, the ongoing interest in characterizing genotype–phenotype relationships has yielded incredible insights into α‐globin gene regulation and its impact on β‐hemoglobinopathies. In this review, we provide a holistic update on α‐globin gene expression stemming from DNA to RNA to protein, as well as epigenetic mechanisms that can impact gene expression and potentially influence phenotypic outcomes. Here, we highlight defined α‐globin targeted strategies and rationalize the use of distinct molecular targets based on the restoration of balanced α/β‐like globin chain synthesis. Considering the therapies that either increase β‐globin synthesis or reactivate γ‐globin gene expression, the modulation of α‐globin chains as a disease modifier for β‐hemoglobinopathies still remains largely uncharted in clinical studies.

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Inclusion of a short hairpin RNA targeting <i>BCL11A</i> into a β-globin expressing vector allows concurrent synthesis of curative adult and fetal hemoglobin

Abstract: Not available.

Cited by 6 publications

References 15 publications

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

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

2021 update on clinical trials in β‐thalassemia

Interplay between α‐thalassemia and β‐hemoglobinopathies: Translating genotype–phenotype relationships into therapies

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