TABLE 1.
Summary of results from pre-clinical studies on antisense oligonucleotides for DUX4 knockdown.
| Study | Chemistry | DUX4 target | Model | DUX4 knockdown (dose) | Other results |
|---|---|---|---|---|---|
| Vanderplanck et al. (2011) | 2′-OMePS | Ex2 SA, Ex3 SA | Primary FSHD myoblasts, differentiated post-treatment | 30% (ex2 SA, 50 nM), 50% (ex3 SA, 10 nM) | Reduced TP53 levels, TRIM43 expression |
| Marsollier et al. (2016) | PMO | Ex3 PAS, down-stream elements | Immortalized FSHD myotubes | 25–52% (50 nM) | Reduced DUX4 downstream gene expression; fusion not affected |
| Chen et al. (2016a) | PMO | Ex2 SA, Ex3 PAS | Primary FSHD myotubes | Not assessed | Reduced DUX4+ nuclei, DUX4 downstream gene expression (only for PAS PMOs); transcriptomic improvements |
| Ex3 PAS | FSHD xenograft mice, 1x e.p. into xenograft, evaluated 2 weeks post-treatment | ∼100% (20 μg) | Reduced DUX4 downstream gene expression | ||
| Ansseau et al. (2017) | 2′-OMePS | Ex2 SA, Ex3 SA | Primary aFSHD and dFSHD myoblasts, differentiated post-treatment | ∼90% (ex2 SA, 50 nM; ex3 SA, 10 nM) | Reduced DUX4+ nuclei; saw improvements in size (in aFSHD but not dFSHD myotubes) |
| Vivo-PMO | Ex3 SA | AAV-DUX4 mice, 1x i.m. TA, evaluated 10 days post-treatment | 30-Fold lower than control vivo-PMO | None | |
| Derenne et al. (2020) | Vivo-PMO | Ex3 SA | DUX4 IMEP mice, 1x i.p., evaluated 1 week post-treatment | Not assessed | 2.5-fold decrease in histological lesion compared to non-treated |
| Lim et al. (2020b) | LNA gapmer | Ex1, Ex3 | Immortalized FSHD myotubes | ∼100% (100 nM) | Reduced DUX4 downstream gene expression; partial transcriptomic restoration; improved muscle cell fusion/size |
| Ex3 | FLExDUX4 mice, 3x i.m., evaluated 1 or 7 days post-treatment | 84% (1 day, 20 μg/i.m.), 70% (7 days, 20 μg/i.m.) | Gapmer uptake observed in and between muscle fibers | ||
| Lim et al. (2020a) | 2′-MOE gapmer | Ex3 | Immortalized FSHD myotubes | ∼100% (100 nM) | Reduced DUX4 downstream gene expression; partial transcriptomic restoration; improved muscle cell fusion/size |
| Ex3 | FLExDUX4 mice, 3x i.m., evaluated 1 day post-treatment | ∼65% (20 μg/i.m.) | None |
2′-OMePS, phosphorothioated 2′-O-methyl RNAs; PMO, phosphorodiamidate morpholino oligomer; LNA, locked nucleic acid; 2′-MOE, 2′-O-methoxyethyl; Ex, exon; SA, splice acceptor; PAS, polyadenylation signal; e.p., electroporation; i.m., intramuscular injection; i.p., intraperitoneal injection; TA, tibialis anterior; AAV, adeno-associated virus; IMEP, intramuscular injection and electroporation of naked plasmid DNA; aFSHD, atrophic FSHD myotubes; dFSHD, disorganized FSHD myotubes.