Hexose enhances oligonucleotide delivery and exon skipping in dystrophin-deficient mdx mice

Hou, Gang; Gu, Ben; Cao, Limin; Gao, Xianjun; Wang, Qingsong; Seow, Yiqi; Zhang, Ning; Wood, Matthew; Yin, HaiFang

doi:10.1038/ncomms10981

Cited by 42 publications

(59 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We anticipate that it will be important to promote cellular uptake of AONs in skeletal muscle cells applied via the circulation. Ongoing efforts focus on various strategies including cell penetrating peptides, exosomes, nanoparticles, and stimulation of ATP-dependent transport using hexose 46, 47, 48, 49, 50. Implementation of enhancers of cellular uptake may form the next generation of AONs that will be tested in vivo as potential targeted therapies of human disease.…”

Section: Discussionmentioning

confidence: 99%

GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle Cells

Wal

Bergsma

Gestel

et al. 2017

Molecular Therapy - Nucleic Acids

View full text Add to dashboard Cite

Pompe disease is a metabolic myopathy caused by deficiency of the acid α-glucosidase (GAA) enzyme and results in progressive wasting of skeletal muscle cells. The c.-32-13T>G (IVS1) GAA variant promotes exon 2 skipping during pre-mRNA splicing and is the most common variant for the childhood/adult disease form. We previously identified antisense oligonucleotides (AONs) that promoted GAA exon 2 inclusion in patient-derived fibroblasts. It was unknown how these AONs would affect GAA splicing in skeletal muscle cells. To test this, we expanded induced pluripotent stem cell (iPSC)-derived myogenic progenitors and differentiated these to multinucleated myotubes. AONs restored splicing in myotubes to a similar extent as in fibroblasts, suggesting that they act by modulating the action of shared splicing regulators. AONs targeted the putative polypyrimidine tract of a cryptic splice acceptor site that was part of a pseudo exon in GAA intron 1. Blocking of the cryptic splice donor of the pseudo exon with AONs likewise promoted GAA exon 2 inclusion. The simultaneous blocking of the cryptic acceptor and cryptic donor sites restored the majority of canonical splicing and alleviated GAA enzyme deficiency. These results highlight the relevance of cryptic splicing in human disease and its potential as therapeutic target for splicing modulation using AONs.

show abstract

Section: Discussionmentioning

confidence: 99%

GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle Cells

Wal

Bergsma

Gestel

et al. 2017

Molecular Therapy - Nucleic Acids

View full text Add to dashboard Cite

show abstract

“…A more potent systemic delivery would have the advantage of targeting multiple skin wounds and also mucosal lesions. Efficient in vivo exon skipping has been achieved after intravenous AON delivery in animal models of Duchenne muscular dystrophy (Alter et al, 2006;Lu et al, 2005;Wu et al, 2008;Yokota et al, 2009), and more recently, a study has shown that a glucose-fructose formulation greatly potentiates the exon skipping activity of intravenously delivered AONs (Han et al, 2016). Furthermore, it would be of interest to compare efficacy and safety profiles of several nucleic acid backbones (phosphorodiamidate morpholino oligomers, peptide nucleic acids, or tricyclo-DNA oligomers, either naked or formulated), because the backbone influences delivery, pharmacology, bio-distribution, toxicology profiles, and ultimately their potential clinical use (Goyenvalle et al, 2015;Saleh et al, 2012).…”

Section: In Vivo Exon Skipping In Col7a1mentioning

confidence: 99%

Targeted Exon Skipping Restores Type VII Collagen Expression and Anchoring Fibril Formation in an In Vivo RDEB Model

Turczynski

Titeux

Tonasso

et al. 2016

Journal of Investigative Dermatology

View full text Add to dashboard Cite

Dystrophic epidermolysis bullosa is a group of orphan genetic skin diseases dominantly or recessively inherited, caused by mutations in COL7A1 encoding type VII collagen, which forms anchoring fibrils. Individuals with recessive dystrophic epidermolysis bullosa develop severe skin and mucosal blistering after mild trauma. The exon skipping strategy consists of modulating splicing of a pre-mRNA to induce skipping of a mutated exon. We have targeted COL7A1 exons 73 and 80, which carry recurrent mutations and whose excision preserves the open reading frame. We first showed the dispensability of these exons for type VII collagen function in vivo. We then showed that transfection of primary recessive dystrophic epidermolysis bullosa keratinocytes and fibroblasts carrying null mutations in exon 73 and/or 80, with 2'-O-methyl antisense oligoribonucleotides, led to efficient ex vivo skipping of these exons (50-95%) and resulted in a significant level (up to 36%) of type VII collagen re-expression. Finally, one or two subcutaneous injections of antisense oligoribonucleotides at doses ranging from 400 μg up to 1 mg restored type VII collagen expression and anchoring fibril formation in vivo in a xenograft model of recessive dystrophic epidermolysis bullosa skin equivalent. This work provides a proof of principle for the treatment of patients with recessive dystrophic epidermolysis bullosa by exon skipping using subcutaneous administration of antisense oligoribonucleotides.

show abstract

“…Notably, a glucose/fructose mixture (GF) and 5% fructose outperformed other analogues in promoting PMO-mediated exon-skipping and dystrophin restoration in mdx mice intramuscularly. 16 Here, we further explore the effect of fructose alone on potentiating other AO chemistries and its systemic potential in mdx mice. Investigation of four different AO chemistries with four top hexose candidates from previous screening in mdx mice indicated that hexose can increase the activities of different AOs.…”

Section: Introductionmentioning

confidence: 99%

Fructose Promotes Uptake and Activity of Oligonucleotides With Different Chemistries in a Context-dependent Manner in mdx Mice

Cao

Hou

Lin

et al. 2016

Molecular Therapy - Nucleic Acids

Self Cite

View full text Add to dashboard Cite

Antisense oligonucleotide (AO)-mediated exon-skipping therapeutics shows great promise in correcting frame-disrupting mutations in the DMD gene for Duchenne muscular dystrophy. However, insufficient systemic delivery limits clinical adoption. Previously, we showed that a glucose/fructose mixture augmented AO delivery to muscle in mdx mice. Here, we evaluated if fructose alone could enhance the activities of AOs with different chemistries in mdx mice. The results demonstrated that fructose improved the potency of AOs tested with the greatest effect on phosphorodiamidate morpholino oligomer (PMO), resulted in a 4.25-fold increase in the number of dystrophin-positive fibres, compared to PMO in saline in mdx mice. Systemic injection of lissamine-labeled PMO with fructose at 25 mg/kg led to increased uptake and elevated dystrophin expression in peripheral muscles, compared to PMO in saline, suggesting that fructose potentiates PMO by enhancing uptake. Repeated intravenous administration of PMO in fructose at 50 mg/kg/week for 3 weeks and 50 mg/kg/month for 5 months restored up to 20% of wild-type dystrophin levels in skeletal muscles with improved functions without detectable toxicity, compared to untreated mdx controls. Collectively, we show that fructose can potentiate AOs of different chemistries in vivo although the effect diminished over repeated administration.

show abstract

Hexose enhances oligonucleotide delivery and exon skipping in dystrophin-deficient mdx mice

Cited by 42 publications

References 34 publications

GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle Cells

GAA Deficiency in Pompe Disease Is Alleviated by Exon Inclusion in iPSC-Derived Skeletal Muscle Cells

Targeted Exon Skipping Restores Type VII Collagen Expression and Anchoring Fibril Formation in an In Vivo RDEB Model

Fructose Promotes Uptake and Activity of Oligonucleotides With Different Chemistries in a Context-dependent Manner in mdx Mice

Contact Info

Product

Resources

About