Generation of iPSC line from a Parkinson patient with PARK7 mutation and CRISPR-edited Gibco human episomal iPSC line to mimic PARK7 mutation

Mazza, Melissa Conti; Beilina, Alexandra; Roosen, Dorien A.; Hauser, David N.; Cookson, Mark

doi:10.1016/j.scr.2021.102506

Cited by 3 publications

(7 citation statements)

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“…Although M17 cells are biochemically similar to the vulnerable neurons in Parkinson’s Disease, they are immortalized and thus have potential alterations in metabolism that could affect glycation. To address this, we also measured glycated products in two distinct neuronal lineages: induced pluripotent stem cell-derived forebrain neurons created from fibroblasts donated by a patient bearing a A111L missense mutation that eliminates steady-state DJ-1 (72) and primary neurons cultured from DJ-1 -/- mice. In both DJ-1-deficient neuronal cultures, glycation products are slightly elevated (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…A commercial iPSC line (ThermoFisher, catalog number A18945) and the DJ-1 knockout iPSC line HT-188 (72) were grown on a layer of MEFs (MTI-GlobalStem, catalog number GSC-6201G) in Essential 8TM media (ThermoFisher, catalog number A1517001) until 100% confluent. Confluent cells were transitioned to N3 media (50% DMEM/F12, 50% Neurobasal with 1x Penicillin-Streptomycin, 0.5x B-27 minus vitamin A, 0.5x N2 supplement, 1x Glutamax, 1x NEAA, 0.055 mM 2-mercaptoethanol and 1μg/ml Insulin) plus 1.5 μM Dorsomorphin (Tocris Bioscience) and 10 μM SB431542 (Stemgent) daily for 11 days.…”

Section: Methodsmentioning

confidence: 99%

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DJ-1 glyoxalase activity makes a modest contribution to cellular defense against methylglyoxal damage in neurons

Mazza

Shuck

Lin

et al. 2022

Preprint

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Human DJ-1 is a cytoprotective protein whose absence causes Parkinson's disease and is also associated with other diseases. DJ-1 has an established role as a redox-regulated protein that defends against oxidative stress and mitochondrial dysfunction. Multiple studies have suggested that DJ-1 is also a protein/nucleic acid deglycase that plays a key role in the repair of glycation damage caused by methylglyoxal (MG), a reactive α-keto aldehyde formed by central metabolism. Contradictory reports suggest that DJ-1 is a glyoxalase but not a deglycase and does not play a major role in glycation defense. Resolving this issue is important for understanding how DJ-1 protects cells against insults that can cause disease. We find that DJ-1 reduces levels of reversible adducts of MG with guanine and cysteine in vitro. The steady-state kinetics of DJ-1 acting on reversible hemithioacetal substrates are fitted adequately with a computational kinetic model that requires only a DJ-1 glyoxalase activity, supporting the conclusion that deglycation is an apparent rather than a true activity of DJ-1. Sensitive and quantitative isotope-dilution mass spectrometry shows that DJ-1 modestly reduces the levels of some irreversible guanine and lysine glycation products in primary and cultured neuronal cell lines and whole mouse brain, consistent with a small but measurable effect on total neuronal glycation burden. However, DJ-1 does not improve cultured cell viability in exogenous MG. In total, our results suggest that DJ-1 is not a deglycase and has only a minor role in protecting neurons against methylglyoxal toxicity.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

DJ-1 glyoxalase activity makes a modest contribution to cellular defense against methylglyoxal damage in neurons

Mazza

Shuck

Lin

et al. 2022

Preprint

Self Cite

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“…Neither of the iPSC lines used is listed as commonly misidentified cell lines by the International Cell Line Authentication Committee. The characterization, karyotyping, sequencing, and confirmation of DJ‐1 ablation for the iPSC lines used in the current study is described in (Mazza et al, 2021).…”

Section: Methodsmentioning

confidence: 99%

“…Line Authentication Committee. The characterization, karyotyping, sequencing, and confirmation of DJ-1 ablation for the iPSC lines used in the current study is described in (Mazza et al, 2021).…”

Section: Murine Primary Neuronal Culturesmentioning

confidence: 99%

“…commercial iPSC line (ThermoFisher, catalog number A18945) and the DJ-1 knockout iPSC line HT-188(Mazza et al, 2021) were grown on a layer of MEFs (MTI-GlobalStem, catalog number GSC-6201G) in Essential 8TM media (ThermoFisher, catalog number A1517001) until 100% confluent. Confluent cells were transitioned to N3 media (50% DMEM/F12, 50% Neurobasal with 1x Penicillin-Streptomycin, 0.5x B-27 minus vitamin A, 0.5x N2 supplement, 1x Glutamax, 1x NEAA, 0.055 mM 2-mercaptoethanol and 1 μg/ml insulin) plus 1.5 μM Dorsomorphin (Tocris Bioscience) and 10 μM SB431542 (Stemgent) daily for 11 days.…”

mentioning

confidence: 99%

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DJ‐1 is not a deglycase and makes a modest contribution to cellular defense against methylglyoxal damage in neurons

Mazza

Shuck

Lin

et al. 2022

Journal of Neurochemistry

Self Cite

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Human DJ-1 is a cytoprotective protein whose absence causes Parkinson's disease and is also associated with other diseases. DJ-1 has an established role as a redoxregulated protein that defends against oxidative stress and mitochondrial dysfunction. Multiple studies have suggested that DJ-1 is also a protein/nucleic acid deglycase that plays a key role in the repair of glycation damage caused by methylglyoxal (MG), a reactive α-keto aldehyde formed by central metabolism. Contradictory reports suggest that DJ-1 is a glyoxalase but not a deglycase and does not play a major role in glycation defense. Resolving this issue is important for understanding how DJ-1 protects cells against insults that can cause disease. We find that DJ-1 reduces levels of reversible adducts of MG with guanine and cysteine in vitro. The steady-state kinetics of DJ-1 acting on reversible hemithioacetal substrates are fitted adequately with a computational kinetic model that requires only a DJ-1 glyoxalase activity, supporting the conclusion that deglycation is an apparent rather than a true activity of DJ-1.Sensitive and quantitative isotope-dilution mass spectrometry shows that DJ-1 modestly reduces the levels of some irreversible guanine and lysine glycation products in primary and cultured neuronal cell lines and whole mouse brain, consistent with a small but measurable effect on total neuronal glycation burden. However, DJ-1 does not improve cultured cell viability in exogenous MG. In total, our results suggest that DJ-1 is not a deglycase and has only a minor role in protecting neurons against methylglyoxal toxicity.

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Human Induced Pluripotent Stem Cell Phenotyping and Preclinical Modeling of Familial Parkinson’s Disease

Kim

Daadi

et al. 2022

Genes

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Parkinson’s disease (PD) is primarily idiopathic and a highly heterogenous neurodegenerative disease with patients experiencing a wide array of motor and non-motor symptoms. A major challenge for understanding susceptibility to PD is to determine the genetic and environmental factors that influence the mechanisms underlying the variations in disease-associated traits. The pathological hallmark of PD is the degeneration of dopaminergic neurons in the substantia nigra pars compacta region of the brain and post-mortem Lewy pathology, which leads to the loss of projecting axons innervating the striatum and to impaired motor and cognitive functions. While the cause of PD is still largely unknown, genome-wide association studies provide evidence that numerous polymorphic variants in various genes contribute to sporadic PD, and 10 to 15% of all cases are linked to some form of hereditary mutations, either autosomal dominant or recessive. Among the most common mutations observed in PD patients are in the genes LRRK2, SNCA, GBA1, PINK1, PRKN, and PARK7/DJ-1. In this review, we cover these PD-related mutations, the use of induced pluripotent stem cells as a disease in a dish model, and genetic animal models to better understand the diversity in the pathogenesis and long-term outcomes seen in PD patients.

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Generation of iPSC line from a Parkinson patient with PARK7 mutation and CRISPR-edited Gibco human episomal iPSC line to mimic PARK7 mutation

Cited by 3 publications

References 4 publications

DJ-1 glyoxalase activity makes a modest contribution to cellular defense against methylglyoxal damage in neurons

DJ-1 glyoxalase activity makes a modest contribution to cellular defense against methylglyoxal damage in neurons

DJ‐1 is not a deglycase and makes a modest contribution to cellular defense against methylglyoxal damage in neurons

Human Induced Pluripotent Stem Cell Phenotyping and Preclinical Modeling of Familial Parkinson’s Disease

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