Engineered U7 snRNA mediates sustained splicing correction in erythroid cells from β-thalassemia/HbE patients

Preedagasamzin, Sarinthip; Nualkaew, Tiwaporn; Pongrujikorn, Tanjitti; Jinawath, Natini; Kole, Ryszard; Fucharoen, Suthat; Jearawiriyapaisarn, Natee; Svasti, Saovaros

doi:10.1016/j.bbrc.2018.03.102

Cited by 10 publications

(13 citation statements)

References 16 publications

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“…Importantly, not only were these models useful for gaining further understanding of the molecular mechanisms involved with the disease, but also as a platform for testing splice switching oligonucleotides [79]. Furthermore, these studies have led to new advances for SSO delivery including the lentiviral vectors that stably express U7 snRNAs carrying splice-switching SSO sequences targeted to the cryptic 5’ splice site recently tested in erythroid cells from patients [80].…”

Section: Figmentioning

confidence: 99%

RNA Splicing and Disease: Animal Models to Therapies

et al. 2019

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Alternative splicing of pre-messenger RNA increases genetic diversity and recent studies estimate that most human multi-exon genes are alternatively spliced. If this process is not highly regulated and accurate, it leads to mis-splicing events which may result in proteins with altered function. A growing body of work has implicated mis-splicing events as contributing to a wide range of diseases including cancer, neurodegenerative diseases, and muscular dystrophies. Understanding the mechanisms that cause aberrant splicing events and how this leads to disease is vital for designing effective therapeutic strategies. This review focuses on advances in therapies targeting splicing and highlights the animal models developed to recapitulate disease phenotypes as a model for testing these therapies.

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Section: Figmentioning

confidence: 99%

RNA Splicing and Disease: Animal Models to Therapies

et al. 2019

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“…Furthermore, 5′ capping, secondary structure and binding with small nuclear ribonucleoprotein (snRNP)–specific proteins render them resistant to nuclease degradation and promises continuous expression. These advantages make SSO-embedded U7 snRNAs attractive, as showed by the successful modulation of pre-mRNA splicing in Duchenne muscular dystrophy 16,18 , spinal muscular atrophy 15 , and β-thalassaemia 10,19 .…”

Section: Discussionmentioning

confidence: 99%

“…Importantly, restoration of correct β-globin mRNA splicing and HbA synthesis by U7.BP + 623 snRNA lentiviral vector was demonstrated in erythropoietic cells derived from β-thalassaemia patients and improvement of erythroid cell differentiation was observed. The potential of this approach was previously demonstrated using a U7 snRNA-based vector targeted to the β E -globin pre-mRNA, which restores correct splicing of β E -globin pre-mRNA and improves the pathology of patients with β E -thalassaemia 19 . Even though complete splicing correction was not achieved in this work, the approximately 10% of correction resulted in phenotypic improvement and should improve the balance of α/β-globin.…”

Section: Discussionmentioning

confidence: 99%

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Restoration of correct βIVS2-654-globin mRNA splicing and HbA production by engineered U7 snRNA in β-thalassaemia/HbE erythroid cells

Nualkaew

Jearawiriyapaisarn

Hongeng

et al. 2019

Sci Rep

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A cytosine to thymine mutation at nucleotide 654 of human β-globin intron 2 (β IVS2-654 ) is one of the most common mutations causing β-thalassaemia in Chinese and Southeast Asians. This mutation results in aberrant β-globin pre-mRNA splicing and prevents synthesis of β-globin protein. Splicing correction using synthetic splice-switching oligonucleotides (SSOs) has been shown to restore expression of the β-globin protein, but to maintain therapeutically relevant levels of β-globin it would require lifelong administration. Here, we demonstrate long-term splicing correction using U7 snRNA lentiviral vectors engineered to target several pre-mRNA splicing elements on the β IVS2-654 -globin pre-mRNA such as cryptic 3′ splice site, aberrant 5′ splice site, cryptic branch point and an exonic splicing enhancer. A double-target engineered U7 snRNAs targeted to the cryptic branch point and an exonic splicing enhancer, U7.BP + 623, was the most effective in a model cell line, HeLa IVS2-654. Moreover, the therapeutic potential of the vector was demonstrated in erythroid progenitor cells derived from β IVS2-654 -thalassaemia/HbE patients, which showed restoration of correctly spliced β-globin mRNA and led to haemoglobin A synthesis, and consequently improved thalassaemic erythroid cell pathology. These results demonstrate proof of concept of using the engineered U7 snRNA lentiviral vector for treatment of β-thalassaemia.

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“… 36 SSOs successfully induced correct splicing and increased normal β-globin production in cell-free extracts, stable cell lines with a mutated β-globin gene, erythroid mononuclear cells from peripheral blood of patients, thalassemic mouse erythroid progenitors, human iPSCs and β-thalassemic mouse model. 36 – 38 , 40 , 41 Among patients with β IVS2−654 thalassemia, erythroid cells, free uptake and syringe load of SSOs into cells increased correct β-globin mRNA up to 77% and HbA production up to 54%. 36 Moreover, in the β IVS2−654 -thalassemia mouse model, intravenous injection of SSOs restored 12% of correct β-globin mRNA in mice peripheral blood or increased 6-fold compared with untreated controls.…”

Section: Nucleic Acid Therapy For β-Thalassemiamentioning

confidence: 99%

<p>Nucleic Acid Therapy for β-Thalassemia</p>

d'Arqom¹

2020

BTT

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β-thalassemia is caused by mutations in the β-globin gene which diminishes or abolishes β-globin chain production. This reduction causes an imbalance of the α/β-globin chain ratio and contributes to the pathogenesis of the disease. Several approaches to reduce the imbalance of the α/β ratio using several nucleic acid-based technologies such as RNAi, lentiviral mediated gene therapy, splice switching oligonucleotides (SSOs) and gene editing technology have been investigated extensively. These approaches aim to reduce excess free α-globin, either by reducing the α-globin chain, restoring β-globin expression and reactivating γ-globin expression, leading a reduced disease severity, treatment necessity, treatment interval, and disease complications, thus, increasing the life quality of the patients and alleviating economic burden. Therefore, nucleic acid-based therapy might become a potential targeted therapy for β-thalassemia.

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Engineered U7 snRNA mediates sustained splicing correction in erythroid cells from β-thalassemia/HbE patients

Cited by 10 publications

References 16 publications

RNA Splicing and Disease: Animal Models to Therapies

RNA Splicing and Disease: Animal Models to Therapies

Restoration of correct βIVS2-654-globin mRNA splicing and HbA production by engineered U7 snRNA in β-thalassaemia/HbE erythroid cells

<p>Nucleic Acid Therapy for β-Thalassemia</p>

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