Dual Myostatin and Dystrophin Exon Skipping by Morpholino Nucleic Acid Oligomers Conjugated to a Cell-penetrating Peptide Is a Promising Therapeutic Strategy for the Treatment of Duchenne Muscular Dystrophy

Background and Objectives: Helicobacter pylori causes several gastrointestinal diseases, including asymptomatic gastritis, chronic peptic ulcer, duodenal ulcer, lymphoma of the mucosa-associated lymphoid tissue (MALT), and gastric adenocarcinoma. In recent years, failure to eradicate H. pylori infections has become an alarming problem for physicians. It is now clear that the current treatment strategies may become ineffective, necessitating the development of innovative antimicrobial compounds as alternative treatments. Materials and Methods: In this experimental study, a cell-penetrating peptide-conjugated peptide nucleic acid (CPP-PNA) was used to target the cagA expression. cagA expression was evaluated using RT-qPCR assay after treatment by the CPPPNA in cell culture and animal model. Additionally, immunogenicity and toxicity of the CPP-PNA were assessed in both cell culture and animal models. Results: Our analysis showed that cagA mRNA levels reduced in H. pylori-infected HT29 cells after treatment with CPPPNA in a dose-dependent manner. Also, cagA expression in bacterial RNA extracted from stomach tissue of mice treated with PNA was reduced compared to that of untreated mice. The expression of inflammatory cytokines also decreased in cells and tissue of H. pylori-infected mice after PNA treatment. The tested CPP-PNA showed no significant adverse effects on cell proliferation of cultured cells and no detectable toxicity and immunogenicity were observed in mice. Conclusion: These results suggest the effectiveness of CPP-PNA in targeting CagA for various research and therapeutic purposes, offering a potential antisense therapy against H. pylori infections.

Download Full-text

Intracellular delivery of a peptide nucleic acid-based hybrid of an autophagy inducing peptide with a cell-penetrating peptide

Organic & Biomolecular Chemistry ◽

10.1039/c9ob02559f ◽

2020 ◽

Vol 18 (10) ◽

pp. 1978-1986

Author(s):

Yoshiyuki Hakata ◽

Suzuka Ishikawa ◽

Takashi Ohtsuki ◽

Masaaki Miyazawa ◽

Mizuki Kitamatsu

Keyword(s):

Nucleic Acid ◽

Nucleic Acids ◽

Peptide Nucleic Acid ◽

Peptide Nucleic Acids ◽

Intracellular Delivery ◽

Cell Penetrating Peptide ◽

Cell Penetrating ◽

A Cell

The conjugate of autophagy-inducing peptide with cell-penetrating peptide formed by hybridization between peptide nucleic acids was delivered into cell and induced effective autophagy.

Download Full-text

Faculty Opinions recommendation of Peptide nucleic acid (PNA) cell penetrating peptide (CPP) conjugates as carriers for cellular delivery of antisense oligomers.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.13939980.15395086 ◽

2012 ◽

Author(s):

Daniel Appella ◽

Ethan Englund

Keyword(s):

Nucleic Acid ◽

Peptide Nucleic Acid ◽

Cell Penetrating Peptide ◽

Cellular Delivery ◽

Cell Penetrating

Download Full-text

Faculty Opinions recommendation of Eteplirsen treatment for Duchenne muscular dystrophy: Exon skipping and dystrophin production.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.733222528.793560200 ◽

2019 ◽

Author(s):

Sue Fletcher

Keyword(s):

Duchenne Muscular Dystrophy ◽

Muscular Dystrophy ◽

Exon Skipping

Download Full-text

Evaluating the potential of novel genetic approaches for the treatment of Duchenne muscular dystrophy

European Journal of Human Genetics ◽

10.1038/s41431-021-00811-2 ◽

2021 ◽

Author(s):

Vratko Himič ◽

Kay E. Davies

Keyword(s):

Duchenne Muscular Dystrophy ◽

Muscular Dystrophy ◽

Structural Integrity ◽

Viral Vector ◽

Exon Skipping ◽

Dystrophin Gene ◽

Pharmacological Treatments ◽

Genetic Sequencing ◽

Reading Frame ◽

Target Effects

AbstractDuchenne muscular dystrophy (DMD) is an X-linked progressive muscle-wasting disorder that is caused by a lack of functional dystrophin, a cytoplasmic protein necessary for the structural integrity of muscle. As variants in the dystrophin gene lead to a disruption of the reading frame, pharmacological treatments have only limited efficacy; there is currently no effective therapy and consequently, a significant unmet clinical need for DMD. Recently, novel genetic approaches have shown real promise in treating DMD, with advancements in the efficacy and tropism of exon skipping and surrogate gene therapy. CRISPR-Cas9 has the potential to be a ‘one-hit’ curative treatment in the coming decade. The current limitations of gene editing, such as off-target effects and immunogenicity, are in fact partly constraints of the delivery method itself, and thus research focus has shifted to improving the viral vector. In order to halt the loss of ambulation, early diagnosis and treatment will be pivotal. In an era where genetic sequencing is increasingly utilised in the clinic, genetic therapies will play a progressively central role in DMD therapy. This review delineates the relative merits of cutting-edge genetic approaches, as well as the challenges that still need to be overcome before they become clinically viable.

Download Full-text

Restoring Protein Expression in Neuromuscular Conditions: A Review Assessing the Current State of Exon Skipping/Inclusion and Gene Therapies for Duchenne Muscular Dystrophy and Spinal Muscular Atrophy

BioDrugs ◽

10.1007/s40259-021-00486-7 ◽

2021 ◽

Author(s):

Omar Sheikh ◽

Toshifumi Yokota

Keyword(s):

Duchenne Muscular Dystrophy ◽

Muscular Dystrophy ◽

Protein Expression ◽

Spinal Muscular Atrophy ◽

Muscular Atrophy ◽

Exon Skipping ◽

Gene Therapies ◽

Current State ◽

Neuromuscular Conditions

Download Full-text

The administration of antisense oligonucleotide golodirsen reduces pathological regeneration in patients with Duchenne muscular dystrophy

Acta Neuropathologica Communications ◽

10.1186/s40478-020-01106-1 ◽

2021 ◽

Vol 9 (1) ◽

Author(s):

Dominic Scaglioni ◽

Francesco Catapano ◽

Matthew Ellis ◽

Silvia Torelli ◽

Darren Chambers ◽

...

Keyword(s):

Duchenne Muscular Dystrophy ◽

Muscular Dystrophy ◽

Antisense Oligonucleotide ◽

De Novo ◽

Exon Skipping ◽

Significant Negative Correlation ◽

Entire Cohort ◽

Associated Proteins ◽

Analysis Platform ◽

Dystrophin Protein

AbstractDuring the last decade, multiple clinical trials for Duchenne muscular dystrophy (DMD) have focused on the induction of dystrophin expression using different strategies. Many of these trials have reported a clear increase in dystrophin protein following treatment. However, the low levels of the induced dystrophin protein have raised questions on its functionality. In our present study, using an unbiased, high-throughput digital image analysis platform, we assessed markers of regeneration and levels of dystrophin associated protein via immunofluorescent analysis of whole muscle sections in 25 DMD boys who received 48-weeks treatment with exon 53 skipping morpholino antisense oligonucleotide (PMO) golodirsen. We demonstrate that the de novo dystrophin induced by exon skipping with PMO golodirsen is capable of conferring a histological benefit in treated patients with an increase in dystrophin associated proteins at the dystrophin positive regions of the sarcolemma in post-treatment biopsies. Although 48 weeks treatment with golodirsen did not result in a significant change in the levels of fetal/developmental myosins for the entire cohort, there was a significant negative correlation between the amount of dystrophin and levels of regeneration observed in different biopsy samples. Our results provide, for the first time, evidence of functionality of induced dystrophin following successful therapeutic intervention in the human.

Download Full-text