CRISPR correction of the Finnish ornithine delta-aminotransferase mutation restores metabolic homeostasis in iPSC from patients with gyrate atrophy

Maldonado, Rocio; Jalil, Sami; Keskinen, Timo; Nieminen, Anni I.; Hyvönen, Mervi E.; Lapatto, Risto; Wartiovaara, Kirmo

doi:10.1016/j.ymgmr.2022.100863

Cited by 4 publications

(7 citation statements)

References 26 publications

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“…A recent ar3cle reported a gene replacement strategy delivering a func3onal copy of OAT to the liver and showed improved func3on and structure of the murine re3na. Another report from our group revealed an increase in the transcrip3on of the gene in mutant cells 14 . Thus, the promise of the novel therapeu3c approach s3ll requires further research to confirm the gene3c and molecular dynamics between the mutant and the replacement sequence, the efficiency in human pa3ents and the actual impact on disease progression.…”

mentioning

confidence: 65%

“…Previous reports suggested that the change c.1205 T>C in OAT destabilises the structure of the protein and leads to its degradation 7 . Accordingly, despite the two-fold increase in mRNA expression of the OAT gene in the mutant cells 14 , they lack the protein. The exact mechanism behind this event remains unexplored.…”

Section: Resultsmentioning

confidence: 96%

“…We generated iPSC from skin biopsies from two non-related voluntary female patients, diagnosed with HOGA in their childhood (ethical permit: #HUS/2754/2019). The reprogramming and characterisation of the cells has been reported and described elsewhere 14 .…”

Section: Methodsmentioning

confidence: 99%

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Finnish gyrate atrophy mutation OAT;c.1205C>T leads to accumulation of intracellular GABA

Sartori-Maldonado,

Wartiovaara

2024

Preprint

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Hyperornithinaemia with gyrate atrophy of choroid and retina (HOGA) is a recessive metabolic disease caused by dysfunction of the ornithine aminotransferase (OAT) gene, leading to ornithine accumulation and a complex metabolic imbalance. This causes retinal degeneration that ultimately evolve to blindness. However, the mechanisms of this degeneration remain unknown. Here, we have conducted untargeted metabolomic analysis in patient-derived induced pluripotent stem cells and their isogenic counterparts. Mutant cells show altered levels of ornithine-related metabolites, including low creatine, proline and glutamate, and elevated arginine and citrulline. The untargeted metabolomics approach revealed changes in the urea cycle and polyamine synthesis pathways with a significant intracellular accumulation of gamma-aminobutyric acid (GABA). Hence, we propose GABA as a key player in the disease pathogenicity, potentially affecting neuronal function in the eye.

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confidence: 65%

Section: Resultsmentioning

confidence: 96%

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Finnish gyrate atrophy mutation OAT;c.1205C>T leads to accumulation of intracellular GABA

Sartori-Maldonado,

Wartiovaara

2024

Preprint

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“…For MIRAS POLG1 genetic correction, electroporation with CRISPR–Cas9 system components was performed as previously described 52 . We used high-efficiency gRNA and a dsDNA donor template including the desired correction along with a novel restriction site for SalI (GˆTCGAC).…”

Section: Methodsmentioning

confidence: 99%

Ancestral allele of DNA polymerase gamma modifies antiviral tolerance

Kang,

Hepojoki,

Maldonado

et al. 2024

Nature

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Mitochondria are critical modulators of antiviral tolerance through the release of mitochondrial RNA and DNA (mtDNA and mtRNA) fragments into the cytoplasm after infection, activating virus sensors and type-I interferon (IFN-I) response1–4. The relevance of these mechanisms for mitochondrial diseases remains understudied. Here we investigated mitochondrial recessive ataxia syndrome (MIRAS), which is caused by a common European founder mutation in DNA polymerase gamma (POLG1)5. Patients homozygous for the MIRAS variant p.W748S show exceptionally variable ages of onset and symptoms5, indicating that unknown modifying factors contribute to disease manifestation. We report that the mtDNA replicase POLG1 has a role in antiviral defence mechanisms to double-stranded DNA and positive-strand RNA virus infections (HSV-1, TBEV and SARS-CoV-2), and its p.W748S variant dampens innate immune responses. Our patient and knock-in mouse data show that p.W748S compromises mtDNA replisome stability, causing mtDNA depletion, aggravated by virus infection. Low mtDNA and mtRNA release into the cytoplasm and a slow IFN response in MIRAS offer viruses an early replicative advantage, leading to an augmented pro-inflammatory response, a subacute loss of GABAergic neurons and liver inflammation and necrosis. A population databank of around 300,000 Finnish individuals6 demonstrates enrichment of immunodeficient traits in carriers of the POLG1 p.W748S mutation. Our evidence suggests that POLG1 defects compromise antiviral tolerance, triggering epilepsy and liver disease. The finding has important implications for the mitochondrial disease spectrum, including epilepsy, ataxia and parkinsonism.

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“…iPSC technology allows for the derivation of patient-specific pluripotent cells that can be differentiated into various cell types, providing a unique model system to study disease mechanisms and test therapeutic interventions in a patient-specific manner. This technology has shown significant potential in drug discovery and personalized medicine, offering insights into the pathophysiology of complex diseases [ 3 , 4 ]. CRISPR-Cas9, on the other hand, provides a robust, precise, and relatively simple method for gene editing that can correct genetic defects at the genome level.…”

Section: Introductionmentioning

confidence: 99%

Induced Pluripotent Stem Cells and CRISPR-Cas9 Innovations for Treating Alpha-1 Antitrypsin Deficiency and Glycogen Storage Diseases

Walsh,

Jin

2024

Cells

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Human induced pluripotent stem cell (iPSC) and CRISPR-Cas9 gene-editing technologies have become powerful tools in disease modeling and treatment. By harnessing recent biotechnological advancements, this review aims to equip researchers and clinicians with a comprehensive and updated understanding of the evolving treatment landscape for metabolic and genetic disorders, highlighting how iPSCs provide a unique platform for detailed pathological modeling and pharmacological testing, driving forward precision medicine and drug discovery. Concurrently, CRISPR-Cas9 offers unprecedented precision in gene correction, presenting potential curative therapies that move beyond symptomatic treatment. Therefore, this review examines the transformative role of iPSC technology and CRISPR-Cas9 gene editing in addressing metabolic and genetic disorders such as alpha-1 antitrypsin deficiency (A1AD) and glycogen storage disease (GSD), which significantly impact liver and pulmonary health and pose substantial challenges in clinical management. In addition, this review discusses significant achievements alongside persistent challenges such as technical limitations, ethical concerns, and regulatory hurdles. Future directions, including innovations in gene-editing accuracy and therapeutic delivery systems, are emphasized for next-generation therapies that leverage the full potential of iPSC and CRISPR-Cas9 technologies.

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CRISPR correction of the Finnish ornithine delta-aminotransferase mutation restores metabolic homeostasis in iPSC from patients with gyrate atrophy

Cited by 4 publications

References 26 publications

Finnish gyrate atrophy mutation OAT;c.1205C>T leads to accumulation of intracellular GABA

Finnish gyrate atrophy mutation OAT;c.1205C>T leads to accumulation of intracellular GABA

Ancestral allele of DNA polymerase gamma modifies antiviral tolerance

Induced Pluripotent Stem Cells and CRISPR-Cas9 Innovations for Treating Alpha-1 Antitrypsin Deficiency and Glycogen Storage Diseases

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