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Updated: Jan 17, 2026
Exon Skipping in Directly Reprogrammed Myotubes Obtained from Human Urine-Derived Cells
Published on: May 7, 2020
Exon Skipping Therapy Restores Ciliary Function in USH2A-Related Retinal Degeneration
Wuyi Li1, Yamei Li1, Yunyu Zhou1
1Department of Ophthalmology, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, People's Republic of China.
Purpose:
This study aimed to evaluate the exon-skipping efficacy and safety of an antisense oligonucleotide (AON) targeting USH2A exon 13 across multiple models, including the Rb1 cell line, humanized USH2A-e13 transgenic mice, and patient-derived retinal organoids. Additionally, we investigated the pathogenic mechanisms of USH2A variants and the therapeutic effects of exon skipping on photoreceptor cilia structure and function.
Methods:
Bioinformatic tools were used to design AONs targeting USH2A exon 13, and their exon-skipping efficiency was assessed at both RNA and protein levels in Rb1 cells. A humanized USH2A-e13 transgenic mouse model was generated via gene editing and received intravitreal AON injections. Retinal distribution, exon-skipping efficiency, and toxicity were evaluated through fundus fluorescence imaging, immunofluorescence staining, droplet digital PCR (ddPCR), Western blot (WB), apoptosis assays, and electroretinography (ERG). Patient-derived induced pluripotent stem cells (iPSCs) were differentiated into retinal organoids and analyzed using transcriptomic profiling, immunofluorescence, ddPCR, WB, apoptosis assays, and transmission electron microscopy (TEM).
Results:
PUMCH-E13 effectively induced exon 13 skipping in the Rb1 cell line, USH2A-e13 mice (44.44% ± 1.61% reduction), and patient-derived retinal organoids (16.4% ± 4.1% reduction). No significant adverse effects were observed through apoptosis assays or ERG. Additionally, treatment with PUMCH-E13 resulted in the restoration of GPR98 and PDZD7 expression within the USH2 complex, alongside the reorganization of microtubule structures in the photoreceptor cilia.
Conclusions:
PUMCH-E13 effectively induces exon 13 skipping in USH2A with low toxicity. Additionally, PUMCH-E13 can promote the restoration of photoreceptor cilia structure in patient-derived retinal organoids, revealing its potential therapeutic mechanism.
