Gene Editing Targeting the DUX4 Polyadenylation Signal: A Therapy for FSHD?

Joubert, Romain; Mariot, Virginie; Charpentier, Maud; Concordet, Jean Paul; Dumonceaux, Julie

doi:10.3390/jpm11010007

Cited by 14 publications

(17 citation statements)

References 33 publications

(51 reference statements)

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“…One obvious advantage over murine transgenic models is that human-to-mouse muscle xenografts are comprised almost exclusively of human tissue and thus provide the best source of mature human muscle, outside of the clinic, to study the specificity and efficacy of drugs designed to treat FSHD. In particular, xenograft models of FSHD are the only setting in which therapeutic approaches targeting the FSHD locus and DUX4 epigenetic or transcriptional regulation [ 157 , 159 , 160 , 161 , 162 , 163 , 164 , 165 ] can be tested in vivo.…”

Section: Animal Models Of Fshdmentioning

confidence: 99%

DUX4 Role in Normal Physiology and in FSHD Muscular Dystrophy

Mocciaro

Runfola

Ghezzi

et al. 2021

Cells

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In the last decade, the sequence-specific transcription factor double homeobox 4 (DUX4) has gone from being an obscure entity to being a key factor in important physiological and pathological processes. We now know that expression of DUX4 is highly regulated and restricted to the early steps of embryonic development, where DUX4 is involved in transcriptional activation of the zygotic genome. While DUX4 is epigenetically silenced in most somatic tissues of healthy humans, its aberrant reactivation is associated with several diseases, including cancer, viral infection and facioscapulohumeral muscular dystrophy (FSHD). DUX4 is also translocated, giving rise to chimeric oncogenic proteins at the basis of sarcoma and leukemia forms. Hence, understanding how DUX4 is regulated and performs its activity could provide relevant information, not only to further our knowledge of human embryonic development regulation, but also to develop therapeutic approaches for the diseases associated with DUX4. Here, we summarize current knowledge on the cellular and molecular processes regulated by DUX4 with a special emphasis on FSHD muscular dystrophy.

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Section: Animal Models Of Fshdmentioning

confidence: 99%

DUX4 Role in Normal Physiology and in FSHD Muscular Dystrophy

Mocciaro

Runfola

Ghezzi

et al. 2021

Cells

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“…Some experiments were conducted using gene editing to try to permanently shut down DUX4 expression by targeting its poly(A) sequence. The results suggest that targeting the DUX4 poly(A) signal to destroy the PAS is possible to some extent [ 25 ], hence a more effective method needs further studies. Single nucleotide exchanges within the hexamer are known to interfere tremendously with mRNA processing and cause a range of diseases.…”

Section: Introductionmentioning

confidence: 99%

Analysis Polyadenylation Signal Usage in Sus scrofa

Zhang

Song

Zhang

et al. 2022

Animals

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RNA polyadenylation is an important step in the messenger RNA (mRNA) maturation process, and the first step is recognizing the polyadenylation signal (PAS). The PAS type and distribution is a key determinant of post-transcriptional mRNA modification and gene expression. However, little is known about PAS usage and alternative polyadenylation (APA) regulation in livestock species. Recently, sequencing technology has enabled the generation of a large amount of sequencing data revealing variation in poly(A) signals and APA regulation in Sus scrofa. We identified 62,491 polyadenylation signals in Sus scrofa using expressed sequence tag (EST) sequences combined with RNA-seq analysis. The composition and usage frequency of polyadenylation signal in Sus scrofa is similar with that of human and mouse. The most highly conserved polyadenylation signals are AAUAAA and AUUAAA, used for over 63.35% of genes. In addition, we also analyzed the U/GU-rich downstream sequence (DSE) element, located downstream of the cleavage site. Our results indicate that APA regulation was widely occurred in Sus scrofa, as in other organisms. Our result was useful for the accurate annotation of RNA 3′ ends in Sus scrofa and the analysis of polyadenylation signal usage in Sus scrofa would give the new insights into the mechanisms of transcriptional regulation.

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“…While several reports have proposed the administration of chemical compounds aimed at RNA interference [15][16][17][18] or blocking signal transduction 19 with promising results to reduce DUX4 activity, the effects of these strategies always depend on continuous treatment due to chemical instability and may force patients to semi-permanent frequent drug administration. Genome editing, however, provides an option to achieve permanent transcriptional repression, but some reports have shown skeptical results after the removal of PAS on 4qA due to stochastic effects and layered of efficiency issues, including delivery, binding, and rewriting as desired 20,21 . The targeting of the DUX4 gene body or promoter is still difficult to control as it is located inside a repetitive D4Z4 unit.…”

Section: Introductionmentioning

confidence: 99%

Hit-and-run silencing of endogenous DUX4 by targeting DNA hypomethylation on D4Z4 repeats in facioscapulohumeral muscular dystrophy

Sasaki-Honda

Jonouchi

Arai

et al. 2022

Preprint

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Facioscapulohumeral muscular dystrophy (FSHD), a progressive skeletal muscle disorder, is epigenetically characterized by DNA hypomethylation of the D4Z4 repeats in the 4q35 region, which enables aberrant DUX4 expression. Sustainable DUX4 suppression is thus a promising therapeutic strategy by which to prevent disease progression, but most of the supposed methods to achieve this depend on the expression of a mediator biochemical entity that would potentially narrow the quality of life of individuals with FSHD in the clinical context. In this study, we report that by applying hit-and-run silencing with dCas9-mediated epigenetic editing targeting DNA hypomethylation on D4Z4 repeats, we could achieve the suppression of endogenous DUX4 in our FSHD patient-derived iPSC model. Notably, DNA methylation was significantly upregulated in FSHD cells and suppression effects were observed for at least two weeks after intervention, which was not the case with transient treatments of typical dCas9-KRAB alone. Off-target analysis showed that despite the potential genome-wide risk for DNA methylation, the impact on the transcriptome was limited. We propose that hit-and-run silencing could be a promising option to prevent disease progression with minimum intervention for individuals with FSHD, motivating further study for clinical development.

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Gene Editing Targeting the DUX4 Polyadenylation Signal: A Therapy for FSHD?

Cited by 14 publications

References 33 publications

DUX4 Role in Normal Physiology and in FSHD Muscular Dystrophy

DUX4 Role in Normal Physiology and in FSHD Muscular Dystrophy

Analysis Polyadenylation Signal Usage in Sus scrofa

Hit-and-run silencing of endogenous DUX4 by targeting DNA hypomethylation on D4Z4 repeats in facioscapulohumeral muscular dystrophy

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