Correction of dystrophia myotonica type 1 pre-mRNA transcripts by artificial trans-splicing

Chen, H Y; Kathirvel, Paramasivam; Yee, Woon Chee; Lai, Poh-San

doi:10.1038/gt.2008.150

Cited by 25 publications

(16 citation statements)

References 26 publications

(40 reference statements)

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“…Trans-splicing has restored factor VIII expression in a hemophilia A knock out mouse model, generating F8 levels above 20% of normal 13 . We and others have shown that RNA trans-splicing can rewrite genes that cause a wide range of diseases including spinal motor atrophy 14 , myotonic dystrophy 15 , epidermolysis bullosa 16,22 , cancer 17,18 and sickle cell anemia 23 . More recently, mRNA and protein expression were corrected in the brains of a mouse model of tau mis-splicing by RNA trans-splicing 24 .…”

Section: Discussionmentioning

confidence: 99%

CD22ΔE12 as a molecular target for corrective repair using RNAtrans-splicing: anti-leukemic activity of a rationally designed RNAtrans-splicing molecule

Uckun

Qazi

et al. 2015

Integrative Biology

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Our recent studies have demonstrated that the CD22 exon 12 deletion (CD22ΔE12) is a characteristic genetic defect of therapy-refractory clones in pediatric B-precursor acute lymphoblastic leukemia (BPL) and implicated the CD22ΔE12 genetic defect in the aggressive biology of relapsed or therapy-refractory pediatric BPL. The purpose of the present study was to further evaluate the biologic significance of the CD22ΔE12 molecular lesion and determine if it could serve as a molecular target for corrective repair using RNA trans-splicing therapy. We show that both pediatric and adult B-lineage lymphoid malignancies are characterized by a very high incidence of the CD22ΔE12 genetic defect. We provide experimental evidence that the correction of the CD22ΔE12 genetic defect in human CD22ΔE12+ BPL cells using a rationally designed CD22 RNA trans-splicing molecule (RTM) caused a pronounced reduction of their clonogenicity. The RTM-mediated correction replaced the downstream mutation-rich segment of Intron 12 and remaining segments of the mutant CD22 pre-mRNA with wildtype CD22 Exons 10-14, thereby preventing the generation of the cis-spliced aberrant CD22ΔE12 product. The anti-leukemic activity of this RTM against BPL xenograft clones derived from CD22ΔE12+ leukemia patients provides the preclinical proof-of-concept that correcting the CD22ΔE12 defect with rationally designed CD22 RTMs may provide the foundation for therapeutic innovations that are needed for successful treatment of high-risk and relapsed BPL patients.

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Section: Discussionmentioning

confidence: 99%

CD22ΔE12 as a molecular target for corrective repair using RNAtrans-splicing: anti-leukemic activity of a rationally designed RNAtrans-splicing molecule

Uckun

Qazi

et al. 2015

Integrative Biology

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“…This is much higher than what has been reported in previous studies using endogenous targets. 5,33 However, despite the high efficiency of Mybpc3 5′- trans -splicing at the mRNA level, the amount of repaired cMyBP-C protein was rather low. This suggests a low efficiency of translation and underlines that mRNA copy number and protein levels do not need to be correlated.…”

Section: Discussionmentioning

confidence: 99%

Repair of Mybpc3 mRNA by 5′-trans-splicing in a Mouse Model of Hypertrophic Cardiomyopathy

Mearini

Stimpel

Krämer

et al. 2013

Molecular Therapy - Nucleic Acids

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RNA trans-splicing has been explored as a therapeutic option for a variety of genetic diseases, but not for cardiac genetic disease. Hypertrophic cardiomyopathy (HCM) is an autosomal-dominant disease, characterized by left ventricular hypertrophy (LVH) and diastolic dysfunction. MYBPC3, encoding cardiac myosin-binding protein C (cMyBP-C) is frequently mutated. We evaluated the 5′-trans-splicing strategy in a mouse model of HCM carrying a Mybpc3 mutation. 5′-trans-splicing was induced between two independently transcribed molecules, the mutant endogenous Mypbc3 pre-mRNA and an engineered pre-trans-splicing molecule (PTM) carrying a FLAG-tagged wild-type (WT) Mybpc3 cDNA sequence. PTMs were packaged into adeno-associated virus (AAV) for transduction of cultured cardiac myocytes and the heart in vivo. Full-length repaired Mybpc3 mRNA represented up to 66% of total Mybpc3 transcripts in cardiac myocytes and 0.14% in the heart. Repaired cMyBP-C protein was detected by immunoprecipitation in cells and in vivo and exhibited correct incorporation into the sarcomere in cardiac myocytes. This study provides (i) the first evidence of successful 5′-trans-splicing in vivo and (ii) proof-of-concept of mRNA repair in the most prevalent cardiac genetic disease. Since current therapeutic options for HCM only alleviate symptoms, these findings open new horizons for causal therapy of the severe forms of the disease.

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“…The defective portion of the target premRNA is removed and replaced with a wild-type cDNA exon contained within the PTM, and a functional gene product is transcribed. The use of SmaRT for the functional correction of a number of disease mutation models has been reported (Mansfield et al, 2000;Liu et al, 2002b;Chao et al, 2003;Tahara et al, 2004;Chen et al, 2009). Using a double transsplicing approach, it has been shown that replacement of the mutated mdx exon 23 was achievable in the mRNA target (Lorain et al, 2010).…”

Section: Spliceosome-mediated Rna Trans-splicingmentioning

confidence: 98%

Genetic Therapeutic Approaches for Duchenne Muscular Dystrophy

2012

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Despite an expansive wealth of research following the discovery of the DMD gene 25 years ago, there is still no curative treatment for Duchenne muscular dystrophy. However, there are currently many promising lines of research, including cell-based therapies and pharmacological reagents to upregulate dystrophin via readthrough of nonsense mutations or by upregulation of the dystrophin homolog utrophin. Here we review genetic-based therapeutic strategies aimed at the amelioration of the DMD phenotype. These include the reintroduction of a copy of the DMD gene into an affected tissue by means of a viral vector; correction of the mutated DMD transcript by antisense oligonucleotide-induced exon skipping to restore the open reading frame; and direct modification of the DMD gene at a chromosomal level through genome editing. All these approaches are discussed in terms of the more recent advances, and the hurdles to be overcome if a comprehensive and effective treatment for DMD is to be found. These hurdles include the need to target all musculature of the body. Therefore any potential treatment would need to be administered systemically. In addition, any treatment needs to have a long-term effect, with the possibility of readministration, while avoiding any potentially detrimental immune response to the vector or transgene.

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Correction of dystrophia myotonica type 1 pre-mRNA transcripts by artificial trans-splicing

Cited by 25 publications

References 26 publications

CD22ΔE12 as a molecular target for corrective repair using RNAtrans-splicing: anti-leukemic activity of a rationally designed RNAtrans-splicing molecule

CD22ΔE12 as a molecular target for corrective repair using RNAtrans-splicing: anti-leukemic activity of a rationally designed RNAtrans-splicing molecule

Repair of Mybpc3 mRNA by 5′-trans-splicing in a Mouse Model of Hypertrophic Cardiomyopathy

Genetic Therapeutic Approaches for Duchenne Muscular Dystrophy

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