Efficient CRISPR/Cas9 nickase-mediated genome editing in an in vitro model of mucopolysaccharidosis IVA

Leal, Andrés Felipe; Alméciga-Díaz, Carlos J.

doi:10.1038/s41434-022-00344-3

Cited by 12 publications

(31 citation statements)

References 69 publications

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“…4 ), whereas for GM01361, GM00958, and GM01259 no significant change in extracellular enzyme activity was observed (data not shown). These data provide further evidence of previous findings that suggest that CRISPR/nCas9 is a suitable genome editing tool for MPS IVA 11 . Also, they show the potential of MLPs as non-viral vectors for the transport and delivery of the CRISPR/nCas9 system, with results similar to those observed for a reference delivery system (i.e., lipofectamine).…”

Section: Resultssupporting

confidence: 86%

“…We evaluated the impact of the CRISPR/nCas9 treatment on the total GAGs and lysosomal mass, the major biomarkers of MPS IVA 11 , 35 . We found a normalization in GAGs levels when CRISPR/nCas9 system was delivered with LP to GM00593 and GM01361 fibroblasts.…”

Section: Resultsmentioning

confidence: 99%

“…Currently, the enzyme replacement therapy (ERT), which consists of the weekly administration of elosulfase alfa (Vimizim), given its low half-life, fails to reach therapeutic effect on the bone manifestations of the disease. Since mutations in the GALNS gene are the primary cause of MPS IVA, gene therapy (GT) continues to be a feature as the most promising strategy 1 , 11 . Although some classical GT-based approaches have been attempted on preclinical assays 12 , 13 , they have not implemented in the clinical practice yet.…”

Section: Introductionmentioning

confidence: 99%

“…In this scenario, we recently reported on the suitability of CRISPR/nCas9-based gene therapy as a potential approach to treat MPS IVA in vitro with undetectable Off-target effects and the recovery of the significant pathological biomarkers to wild-type levels 11 . CRISPR/Cas9-based systems have also been used for other lysosomal storage disorders (LSD), with encouraging results 19 – 27 .…”

Section: Introductionmentioning

confidence: 99%

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Delivery and assessment of a CRISPR/nCas9-based genome editing system on in vitro models of mucopolysaccharidoses IVA assisted by magnetite-based nanoparticles

Leal

Cifuentes

Torres

et al. 2022

Sci Rep

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Mucopolysaccharidosis IV A (MPS IVA) is a lysosomal disorder caused by mutations in the GALNS gene. Consequently, the glycosaminoglycans (GAGs) keratan sulfate and chondroitin 6-sulfate accumulate in the lysosomal lumen. Although enzyme replacement therapy has shown essential advantages for the patients, several challenges remain to overcome, such as the limited impact on the bone lesion and recovery of oxidative profile. Recently, we validated a CRISPR/nCas9-based gene therapy with promising results in an in vitro MPS IVA model. In this study, we have expanded the use of this CRISPR/nCas9 system to several MPS IVA fibroblasts carrying different GALNS mutations. Considering the latent need to develop more safety vectors for gene therapy, we co-delivered the CRISPR/nCas9 system with a novel non-viral vector based on magnetoliposomes (MLPs). We found that the CRISPR/nCas9 treatment led to an increase in enzyme activity between 5 and 88% of wild-type levels, as well as a reduction in GAGs accumulation, lysosomal mass, and mitochondrial-dependent oxidative stress, in a mutation-dependent manner. Noteworthy, MLPs allowed to obtain similar results to those observed with the conventional transfection agent lipofectamine. Overall, these results confirmed the potential of CRISPR/nCas9 as a genome editing tool for treating MPS IVA. We also demonstrated the potential use of MLPs as a novel delivery system for CRISPR/nCas9-based therapies.

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Section: Resultssupporting

confidence: 86%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Delivery and assessment of a CRISPR/nCas9-based genome editing system on in vitro models of mucopolysaccharidoses IVA assisted by magnetite-based nanoparticles

Leal

Cifuentes

Torres

et al. 2022

Sci Rep

Self Cite

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“…In another study, using CRISPR-Cas9, the authors corrected murine CD34 + hematopoietic stem cells targeting the lysosomal enzyme iduronidase into the CCR5 locus, leading to iduronidase secretion and improved biochemical and phenotypic abnormalities in an MPS I model [ 75 ]. Similarly, CRISPR-Cas9 insertion into the AAVS1 locus generated a 40% increment in GALNS activity, while lysosomal mass, total GAGs, and oxidative stress were normalized [ 76 ]. Miki et al used the ex vivo gene-editing therapy using induced pluripotent stem cell (iPSC) and CRISPR/Cas9 technologies in an MPS Type 1 disease mouse model.…”

Section: Mucopolysaccharidosismentioning

confidence: 99%

Novel Gene-Correction-Based Therapeutic Modalities for Monogenic Liver Disorders

et al. 2022

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The majority of monogenic liver diseases are autosomal recessive disorders, with few being sex-related or co-dominant. Although orthotopic liver transplantation (LT) is currently the sole therapeutic option for end-stage patients, such an invasive surgical approach is severely restricted by the lack of donors and post-transplant complications, mainly associated with life-long immunosuppressive regimens. Therefore, the last decade has witnessed efforts for innovative cellular or gene-based therapeutic strategies. Gene therapy is a promising approach for treatment of many hereditary disorders, such as monogenic inborn errors. The liver is an organ characterized by unique features, making it an attractive target for in vivo and ex vivo gene transfer. The current genetic approaches for hereditary liver diseases are mediated by viral or non-viral vectors, with promising results generated by gene-editing tools, such as CRISPR-Cas9 technology. Despite massive progress in experimental gene-correction technologies, limitations in validated approaches for monogenic liver disorders have encouraged researchers to refine promising gene therapy protocols. Herein, we highlighted the most common monogenetic liver disorders, followed by proposed genetic engineering approaches, offered as promising therapeutic modalities.

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Genome Editing Tools for Lysosomal Storage Disorders

Gonzalez

Nader

Siebert

et al. 2023

Advances in Experimental Medicine and Biology

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Efficient CRISPR/Cas9 nickase-mediated genome editing in an in vitro model of mucopolysaccharidosis IVA

Cited by 12 publications

References 69 publications

Delivery and assessment of a CRISPR/nCas9-based genome editing system on in vitro models of mucopolysaccharidoses IVA assisted by magnetite-based nanoparticles

Delivery and assessment of a CRISPR/nCas9-based genome editing system on in vitro models of mucopolysaccharidoses IVA assisted by magnetite-based nanoparticles

Novel Gene-Correction-Based Therapeutic Modalities for Monogenic Liver Disorders

Genome Editing Tools for Lysosomal Storage Disorders

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