Antisense oligonucleotide therapy rescues aggresome formation in a novel spinocerebellar ataxia type 3 human embryonic stem cell line

Moore, Lauren; Keller, Laura; Bushart, David D.; Delatorre, Rodrigo G.; Li, Duojia; McLoughlin, Hayley S.; Costa, Maria do Carmo; Shakkottai, Vikram G.; Smith, Gary D.; Paulson, Henry L.

doi:10.1016/j.scr.2019.101504

Cited by 37 publications

(26 citation statements)

References 65 publications

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“…In polyQ disease stem cell models such as SCA3, previous studies have shown that polyQ aggregations and neurotoxic effects can only be detected under exogenous stressor stimulation or genetic manipulation [ 42 , 67 – 69 ]. The afore-mentioned results confirmed the formation of the aggregates in SCA3 cell models under normal stress-free culture conditions, which may relate to its neurotoxicity, similar to previous studies [ 70 , 71 ]. Moreover, In the absence of exogenous stressors, the human cell model showing endogenous expression of m ATXN3 aggregations will be a good tool to help us further understand the role of endogenous ATXN3 misfolding and aggregation in the pathogenesis of SCA3.…”

Section: Discussionsupporting

confidence: 91%

CRISPR/Cas9 mediated gene correction ameliorates abnormal phenotypes in spinocerebellar ataxia type 3 patient-derived induced pluripotent stem cells

Wang

Peng

et al. 2021

Transl Psychiatry

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Spinocerebellar ataxia type 3/Machado–Joseph disease (SCA3/MJD) is a progressive autosomal dominant neurodegenerative disease caused by abnormal CAG repeats in the exon 10 of ATXN3. The accumulation of the mutant ataxin-3 proteins carrying expanded polyglutamine (polyQ) leads to selective degeneration of neurons. Since the pathogenesis of SCA3 has not been fully elucidated, and no effective therapies have been identified, it is crucial to investigate the pathogenesis and seek new therapeutic strategies of SCA3. Induced pluripotent stem cells (iPSCs) can be used as the ideal cell model for the molecular pathogenesis of polyQ diseases. Abnormal CAG expansions mediated by CRISPR/Cas9 genome engineering technologies have shown promising potential for the treatment of polyQ diseases, including SCA3. In this study, SCA3-iPSCs can be corrected by the replacement of the abnormal CAG expansions (74 CAG) with normal repeats (17 CAG) using CRISPR/Cas9-mediated homologous recombination (HR) strategy. Besides, corrected SCA3-iPSCs retained pluripotent and normal karyotype, which can be differentiated into a neural stem cell (NSCs) and neuronal cells, and maintained electrophysiological characteristics. The expression of differentiation markers and electrophysiological characteristics were similar among the neuronal differentiation from normal control iPSCs (Ctrl-iPSCs), SCA3-iPSCs, and isogenic control SCA3-iPSCs. Furthermore, this study proved that the phenotypic abnormalities in SCA3 neurons, including aggregated IC2-polyQ protein, decreased mitochondrial membrane potential (MMP) and glutathione expressions, increased reactive oxygen species (ROS), intracellular Ca2+ concentrations, and lipid peroxidase malondialdehyde (MDA) levels, all were rescued in the corrected SCA3-NCs. For the first time, this study demonstrated the feasibility of CRISPR/Cas9-mediated HR strategy to precisely repair SCA3-iPSCs, and reverse the corresponding abnormal disease phenotypes. In addition, the importance of genetic control using CRISPR/Cas9-mediated iPSCs for disease modeling. Our work may contribute to providing a potential ideal model for molecular mechanism research and autologous stem cell therapy of SCA3 or other polyQ diseases, and offer a good gene therapy strategy for future treatment.

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Section: Discussionsupporting

confidence: 91%

CRISPR/Cas9 mediated gene correction ameliorates abnormal phenotypes in spinocerebellar ataxia type 3 patient-derived induced pluripotent stem cells

Wang

Peng

et al. 2021

Transl Psychiatry

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“…S8). The autophagy-linked ubiquitin-binding shuttle protein p62 (SQSTM1) is known to co-localize with both wt and mutATXN3 proteins and promote perinuclear aggresome formation [33,74,[77][78][79]. The non-allele-specific knockdown of ATXN3 in iPSCs did not show significant differences in p62-aggresome formation from untreated SCA3 iPSCs or normal WTC11 iPSCs (Fig.…”

Section: Lna8-17dz9 Targets Both Normal and Expanded Atxn3 Alleles Without Affecting P62-aggresome Levels In Sca3 Ipscsmentioning

confidence: 95%

“…Significant progresses have been made through CRISPR-Cas genome editing and antisense oligonucleotide (ASO) therapies in many polyQ diseases, especially in HD, SCA2, and SCA3 [27][28][29][30][31][32][33][34][35][36][37][38]. Efforts towards engineering high-fidelity Cas proteins with various controller modules, split Cas proteins for efficient viral packaging, and non-viral delivery approaches are underway to improve precision and safety [39][40][41][42].…”

Section: Introductionmentioning

confidence: 99%

“…Efforts towards engineering high-fidelity Cas proteins with various controller modules, split Cas proteins for efficient viral packaging, and non-viral delivery approaches are underway to improve precision and safety [39][40][41][42]. ASOs have been successfully tested in HD, SCA1, SCA2, SCA3, and SCA7 cells or mouse models and with HD ASO entering the Phase III clinical trial [30,33,34,36,37,[43][44][45]. The efficacy of ASO may vary from case to case and may largely depend on RNase H activity, accessibility of target sites, design chemistry, and bioavailability in target tissue.…”

Section: Introductionmentioning

confidence: 99%

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DNAzyme Cleavage of CAG Repeat RNA in Polyglutamine Diseases

et al. 2021

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CAG repeat expansion is the genetic cause of nine incurable polyglutamine (polyQ) diseases with neurodegenerative features. Silencing repeat RNA holds great therapeutic value. Here, we developed a repeat-based RNA-cleaving DNAzyme that catalyzes the destruction of expanded CAG repeat RNA of six polyQ diseases with high potency. DNAzyme preferentially cleaved the expanded allele in spinocerebellar ataxia type 1 (SCA1) cells. While cleavage was non-allele-specific for spinocerebellar ataxia type 3 (SCA3) cells, treatment of DNAzyme leads to improved cell viability without affecting mitochondrial metabolism or p62-dependent aggresome formation. DNAzyme appears to be stable in mouse brain for at least 1 month, and an intermediate dosage of DNAzyme in a SCA3 mouse model leads to a significant reduction of high molecular weight ATXN3 proteins. Our data suggest that DNAzyme is an effective RNA silencing molecule for potential treatment of multiple polyQ diseases.

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“…Especially AONs bear great therapeutic potential as they can be diseaseand allele-specific, are relatively easy to administer, and are efficiently taken up by brain cells with a stability for months [124]. Hitherto, only SCA36 hiPSC-derived motor neurons have been treated with AONs, but several animal and cell models for SCA1, SCA2, SCA3 and SCA7 have been highly instrumental to test different AONs with variable success rates [125][126][127][128][129][130][131][132][133][134]. Moreover, the rapid development of genome editing strategies by CRISPR/Cas makes it possible to correct causative genetic variation, which is of ultimate importance in personalized medicine.…”

Section: Therapeutic Perspectivesmentioning

confidence: 99%

Human Induced Pluripotent Stem Cell-Based Modelling of Spinocerebellar Ataxias

Hommersom

Buijsen

Roon-Mom

et al. 2021

Stem Cell Rev and Rep

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Dominant spinocerebellar ataxias (SCAs) constitute a large group of phenotypically and genetically heterogeneous disorders that mainly present with dysfunction of the cerebellum as their main hallmark. Although animal and cell models have been highly instrumental for our current insight into the underlying disease mechanisms of these neurodegenerative disorders, they do not offer the full human genetic and physiological context. The advent of human induced pluripotent stem cells (hiPSCs) and protocols to differentiate these into essentially every cell type allows us to closely model SCAs in a human context. In this review, we systematically summarize recent findings from studies using hiPSC-based modelling of SCAs, and discuss what knowledge has been gained from these studies. We conclude that hiPSC-based models are a powerful tool for modelling SCAs as they contributed to new mechanistic insights and have the potential to serve the development of genetic therapies. However, the use of standardized methods and multiple clones of isogenic lines are essential to increase validity and reproducibility of the insights gained. Graphical Abstract

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Antisense oligonucleotide therapy rescues aggresome formation in a novel spinocerebellar ataxia type 3 human embryonic stem cell line

Cited by 37 publications

References 65 publications

CRISPR/Cas9 mediated gene correction ameliorates abnormal phenotypes in spinocerebellar ataxia type 3 patient-derived induced pluripotent stem cells

CRISPR/Cas9 mediated gene correction ameliorates abnormal phenotypes in spinocerebellar ataxia type 3 patient-derived induced pluripotent stem cells

DNAzyme Cleavage of CAG Repeat RNA in Polyglutamine Diseases

Human Induced Pluripotent Stem Cell-Based Modelling of Spinocerebellar Ataxias

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