Development of DG9 peptide-conjugated single- and multi-exon skipping therapies for the treatment of Duchenne muscular dystrophy

Lim, Kenji Rowel Q.; Woo, Stanley; Melo, Dyanna; Huang, Yiqing; Dzierlega, Kasia; Shah, Nur Ahad; Aslesh, Tejal; Roshmi, Rohini Roy; Echigoya, Yusuke; Maruyama, Rika; Moulton, Hong M.; Yokota, Toshifumi

doi:10.1073/pnas.2112546119

Cited by 29 publications

(22 citation statements)

References 37 publications

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“…We employed a cell-penetrating peptide DG9 derived from the PTD of the human Hph-1 transcription factor, since fusion proteins containing the Hph-1 domain are known to be delivered efficiently to a wide variety of tissues, including the heart (35). We recently demonstrated successful single and multi-exon skipping using DG9 conjugated to a different PMO cocktail targeting exons 45-55 in the dystrophin gene in vivo for the treatment of Duchenne muscular dystrophy (DMD) (54). Similar to our DMD findings, DG9-PMO treatment resulted in higher SMN expression in vivo compared to unconjugated PMO and MOE (Figure 2).…”

Section: Discussionmentioning

confidence: 99%

DG9-conjugated morpholino rescues phenotype in SMA mice by reaching the CNS via a subcutaneous administration

et al. 2023

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Antisense oligonucleotide (AO)-mediated therapy is a promising strategy to treat several neurological diseases including spinal muscular atrophy (SMA). However, limited delivery to the central nervous system (CNS) with AOs administered intravenously or subcutaneously is a major challenge. Here we demonstrate a single subcutaneous administration of cell-penetrating peptide DG9 conjugated to an AO called phosphorodiamidate morpholino oligomers (PMOs) reaches the CNS and significantly prolonged the median survival compared to unconjugated PMO and R6G-PMO in a severe SMA mouse model. Treated mice exhibited significantly higher expression of full-length SMN2 expression (FL-SMN2) in both the CNS and systemic tissues compared to non-treated and unmodified AO-treated mice. The treatment ameliorated the atrophic musculature and improved breathing function accompanied by improved muscle strength and innervation at the neuromuscular junction with no signs of apparent toxicity. We also demonstrated DG9-conjugated PMO localizes in nuclei in the spinal cord and brain after subcutaneous injections. Our data identify DG9 peptide conjugation as a powerful way to improve the efficacy of AO-mediated splice modulation. Finally, DG9-PMO is a promising therapeutic option to treat SMA and other neurological diseases, overcoming the necessity for intrathecal injections and treating body-wide tissues without apparent toxicity.

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Section: Discussionmentioning

confidence: 99%

DG9-conjugated morpholino rescues phenotype in SMA mice by reaching the CNS via a subcutaneous administration

et al. 2023

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“…Exonic duplications and deletions within DMD are the main pathogenic variants ( 44 ), which has gained special interest as a new therapeutic method for DMD ( 45 ). It is reported that DMD exons 45-55 deletion has been postulated as a model that could treat up to 60% of DMD patients ( 46 ).…”

Section: Discussionmentioning

confidence: 99%

Identification of immune-related features involved in Duchenne muscular dystrophy: A bidirectional transcriptome and proteome-driven analysis

Dong

et al. 2022

Front. Immunol.

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BackgroundWe aimed to investigate the biological mechanism and feature genes of Duchenne muscular dystrophy (DMD) by multi-omics and experimental verification strategy.MethodsWe integrated the transcriptomic and proteomic methods to find the differentially expressed mRNAs (DEMs) and proteins (DEPs) between DMD and Control groups. Weighted gene co-expression network analysis (WGCNA) was then used to identify modules of highly correlated genes and hub genes. In the following steps, the immune and stromal cells infiltrations were accomplished by xCELL algorithm. Furthermore, TF and miRNA prediction were performed with Networkanalyst. ELISA, western blot and external datasets were performed to verify the key proteins/mRNAs in DMD patient and mouse. Finally, a nomogram model was established based on the potential biomarkers.Results4515 DEMs and 56 DEPs were obtained from the transcriptomic and proteomic study respectively. 14 common genes were identified, which is enriched in muscle contraction and inflammation-related pathways. Meanwhile, we observed 33 significant differences in the infiltration of cells in DMD. Afterwards, a total of 22 miRNAs and 23 TF genes interacted with the common genes, including TFAP2C, MAX, MYC, NFKB1, RELA, hsa-miR-1255a, hsa-miR-130a, hsa-miR-130b, hsa-miR-152, and hsa-miR-17. In addition, three genes (ATP6AP2, CTSS, and VIM) showed excellent diagnostic performance on discriminating DMD in GSE1004, GSE3307, GSE6011 and GSE38417 datasets (all AUC > 0.8), which is validated in patients (10 DMD vs. 10 controls), DMD with exon 55 mutations, mdx mouse, and nomogram model.ConclusionTaken together, ATP6AP2, CTSS, and VIM play important roles in the inflammatory response in DMD, which may serve as diagnostic biomarkers and therapeutic targets.

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“…This method improves the applicability of antisense therapy for DMD, and a cocktail skipping exons 45-55 has been validated in vitro and in vivo which could be applicable to nearly 50% of all DMD patients [69][70][71]. Furthermore, this research has demonstrated that spontaneous skipping can occur during multi-exon skipping which reduces the number of AONs required in the cocktail, reducing treatment complexity and cost [72]. At this time, the development of multi-exon skipping cocktails is limited by regulatory hurdles, as each AON in the cocktail must undergo separate clinical approval [24].…”

Section: Dyne-251mentioning

confidence: 94%

Recent Trends in Antisense Therapies for Duchenne muscular dystrophy

Wilton-Clark¹,

Yokota²

2023

Preprint

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Duchenne muscular dystrophy (DMD) is a debilitating and fatal genetic disease affecting 1/3500 boys globally, characterized by progressive muscle breakdown and eventual death with an average lifespan in the mid-late twenties. While no cure yet exists for DMD, gene and antisense therapies have been heavily explored in recent years to better treat this disease. Four antisense therapies have received conditional FDA approval, and many more exist in varying stages of clinical trials. These upcoming therapies often utilize novel drug chemistries to address limitations of existing therapies, and their development could herald the next generation of antisense therapy. This review article aims to summarize the current state of development for antisense-based therapies for the treatment of Duchenne muscular dystrophy, exploring candidates designed for both exon skipping and gene knockdown.

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Development of DG9 peptide-conjugated single- and multi-exon skipping therapies for the treatment of Duchenne muscular dystrophy

Cited by 29 publications

References 37 publications

DG9-conjugated morpholino rescues phenotype in SMA mice by reaching the CNS via a subcutaneous administration

DG9-conjugated morpholino rescues phenotype in SMA mice by reaching the CNS via a subcutaneous administration

Identification of immune-related features involved in Duchenne muscular dystrophy: A bidirectional transcriptome and proteome-driven analysis

Recent Trends in Antisense Therapies for Duchenne muscular dystrophy

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