Rescue of imprinted genes by epigenome editing in human cellular models of Prader-Willi syndrome
Akisa Nemoto1,2,3, Kent Imaizumi1,2,4,5, Fuyuki Miya6
1Department of Physiology, Keio University School of Medicine, Tokyo, Japan.
Insights
Prader-Willi syndrome (PWS) is a genomic imprinting disorder. CRISPR epigenome editing successfully reactivated silenced genes in patient cells, offering a potential therapeutic strategy for PWS and similar conditions.
Area of Science:
- Genetics
- Epigenetics
- Developmental Biology
Background:
- Prader-Willi syndrome (PWS) is a complex genomic imprinting disorder.
- It stems from the loss of function of paternal chromosome 15q11-13, leading to hypothalamic dysfunction and associated symptoms.
- PWS patients exhibit absent expression of paternally expressed genes (PEGs) from the maternal allele due to epigenetic silencing.
Purpose of the Study:
- To investigate the therapeutic potential of CRISPR-based epigenome editing for PWS.
- To modulate the DNA methylation status of the PWS imprinting control region (PWS-ICR) in patient-derived cells.
- To assess the restoration of PEG expression and epigenetic patterns following editing.
Main Methods:
- Utilized a CRISPR-based epigenome editing system to target the PWS-ICR in induced pluripotent stem cells (iPSCs) from PWS patients.
- Performed demethylation of the PWS-ICR to reactivate silenced maternal alleles.
- Differentiated edited iPSCs into hypothalamic organoids and conducted single-cell transcriptomic analysis.
Main Results:
- Successful demethylation of the PWS-ICR restored PEG expression from the maternal allele.
- Epigenome editing reorganized methylation patterns in other PWS-associated imprinted regions.
- Corrected epigenetic patterns and PEG expression were maintained after differentiation into hypothalamic organoids.
- Single-cell transcriptomics revealed partial restoration of transcriptomic dysregulation in edited organoids.
Conclusions:
- CRISPR-based epigenome editing is a viable therapeutic strategy for Prader-Willi syndrome.
- This approach can restore gene expression and correct epigenetic abnormalities in PWS.
- Epigenome editing holds promise for treating PWS and other imprinting disorders.
Abstract:
Prader-Willi syndrome (PWS) is a genomic imprinting disorder caused by the loss of function of the paternal chromosome 15q11-13, resulting in a spectrum of symptoms associated with hypothalamic dysfunction. PWS patients lack the expression of paternally expressed genes (PEGs) in the 15q11-13 locus but possess an epigenetically silenced set of these genes in the maternal allele. Thus, activation of these silenced genes can serve as a therapeutic target for PWS. Here, we leverage CRISPR-based epigenome editing system to modulate the DNA methylation status of the PWS imprinting control region (PWS-ICR) in induced pluripotent stem cells (iPSCs) derived from PWS patients. Successful demethylation in the PWS-ICR restores the PEG expression from the maternal allele and reorganizes the methylation patterns in other PWS-associated imprinted regions beyond the PWS-ICR. Remarkably, these corrected epigenomic patterns and PEG expression are maintained following the differentiation of these cells into hypothalamic organoids. Finally, the single-cell transcriptomic analysis of epigenome-edited organoids demonstrates a partial restoration of the transcriptomic dysregulation observed in PWS. This study highlights the utility of epigenome editing technology as a therapeutic approach in addressing PWS and potentially other imprinting disorders.
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