Human neuronal networks on micro-electrode arrays are a highly robust tool to study disease-specific genotype-phenotype correlations in vitro

Stem Cell Reports, Год журнала: 2021, Номер 16(9), С. 2182 - 2196

Опубликована: Июль 29, 2021

Micro-electrode arrays (MEAs) are increasingly used to characterize neuronal network activity of human induced pluripotent stem cell (hiPSC)-derived neurons. Despite their gain in popularity, MEA recordings from hiPSC-derived networks not always full potential respect experimental design, execution, and data analysis. Therefore, we benchmarked the robustness MEA-derived patterns ten healthy individual control lines, uncover comparable phenotypes. To achieve standardization, provide recommendations on design With such MEAs can be as a reliable platform distinguish (disease-specific) In conclusion, show that powerful robust tool functional phenotypes networks, an important resource advance hiPSC field toward use for disease phenotyping drug discovery.

Язык: Английский

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Imbalanced autophagy causes synaptic deficits in a human model for neurodevelopmental disorders

Autophagy, Год журнала: 2021, Номер 18(2), С. 423 - 442

Опубликована: Июль 21, 2021

Macroautophagy (hereafter referred to as autophagy) is a finely tuned process of programmed degradation and recycling proteins cellular components, which crucial in neuronal function synaptic integrity. Mounting evidence implicates chromatin remodeling fine-tuning autophagy pathways. However, this epigenetic regulation poorly understood neurons. Here, we investigate the role KANSL1, member nonspecific lethal complex, acetylates histone H4 on lysine 16 (H4K16ac) facilitate transcriptional activation. Loss-of-function KANSL1 strongly associated with neurodevelopmental disorder Koolen-de Vries Syndrome (KdVS). Starting from KANSL1-deficient human induced-pluripotent stem cells, both KdVS patients genome-edited lines, identified SOD1 (superoxide dismutase 1), an antioxidant enzyme, be significantly decreased, leading subsequent increase oxidative stress autophagosome accumulation. In neurons, accumulation at excitatory synapses resulted reduced density, GRIA/AMPA receptor-mediated transmission impaired network activity. Furthermore, found that increased stress-mediated leads MTOR activation decreased lysosome function, further preventing clearing autophagosomes. Finally, by pharmacologically reducing stress, could rescue aberrant formation well activity Our findings thus point toward important relation between stress-induced synapse demonstrate importance H4K16ac-mediated changes structure balance reactive oxygen species- MTOR-dependent autophagy.Abbreviations: APO: apocynin; ATG: related; BAF: bafilomycin A1; BSO: buthionine sulfoximine; CV: coefficient variation; DIV: days vitro; H4K16ac: 4 acetylation; iPSC: cell; KANSL1: KAT8 regulatory NSL complex subunit 1; KdVS: Syndrome; LAMP1: lysosomal membrane protein MAP1LC3/LC3: microtubule 1 light chain 3; MEA: micro-electrode array; MTOR: mechanistic target rapamycin kinase; complex: complex; 8-oxo-dG: 8-hydroxydesoxyguanosine; RAP: rapamycin; ROS: species; sEPSCs: spontaneous postsynaptic currents; SOD1: superoxide SQSTM1/p62: sequestosome SYN: synapsin; WRT: wortmannin.

Язык: Английский

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iPSC models of mitochondrial diseases

Neurobiology of Disease, Год журнала: 2025, Номер unknown, С. 106822 - 106822

Опубликована: Янв. 1, 2025

Язык: Английский

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A MYT1L syndrome mouse model recapitulates patient phenotypes and reveals altered brain development due to disrupted neuronal maturation

Neuron, Год журнала: 2021, Номер 109(23), С. 3775 - 3792.e14

Опубликована: Окт. 5, 2021

Язык: Английский

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Pathogenic mitochondrial DNA 3243A>G mutation: From genetics to phenotype

Frontiers in Genetics, Год журнала: 2022, Номер 13

Опубликована: Окт. 6, 2022

The mitochondrial DNA (mtDNA) m.3243A>G mutation is one of the most common pathogenic mtDNA variants, showing complex genetics, molecular mechanisms, and phenotypes. In recent years, prevention mtDNA-related diseases has trended toward precision medicine strategies, such as preimplantation genetic diagnosis (PGD) replacement therapy (MRT). These techniques are set to allow birth healthy children, but clinical implementation relies on thorough insights into genetics. genotype phenotype vary greatly from mother offspring, which compromises counseling for disease. This review first systematically elaborate characteristics mutation, genetics relationship between them, well related influencing factors potential strategies preventing perceptions will provide clarity clinicians providing patients.

Язык: Английский

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The potential of in vitro neuronal networks cultured on micro electrode arrays for biomedical research

Progress in Biomedical Engineering, Год журнала: 2023, Номер 5(3), С. 032002 - 032002

Опубликована: Апрель 18, 2023

Abstract In vitro neuronal models have become an important tool to study healthy and diseased circuits. The growing interest of neuroscientists explore the dynamics systems increasing need observe, measure manipulate not only single neurons but populations cells pushed for technological advancement. this sense, micro-electrode arrays (MEAs) emerged as a promising technique, made cell culture dishes with embedded micro-electrodes allowing non-invasive relatively simple measurement activity cultures at network level. past decade, MEAs popularity has rapidly grown. MEA devices been extensively used mainly derived from rodents. Rodent on employed investigate physiological mechanisms, effect chemicals in neurotoxicity screenings, model electrophysiological phenotype networks different pathological conditions. With advancements human induced pluripotent stem (hiPSCs) technology, differentiation adult donors became possible. hiPSCs-derived develop patient-specific platforms characterize pathophysiological test drugs, paving way towards personalized medicine. review, we first describe technology information that can be obtained recordings. Then, give overview studies which combination (i.e. rodent 2D three-dimensional (3D) cultures, organotypic brain slices, 3D organoids) biomedical research, including physiology studies, disease modeling, drug testing. We end by discussing potential, challenges future perspectives providing some guidance choice device, experimental design, data analysis reporting scientific publications.

Язык: Английский

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Modeling mitochondrial DNA diseases: from base editing to pluripotent stem‐cell‐derived organoids

EMBO Reports, Год журнала: 2023, Номер 24(4)

Опубликована: Март 6, 2023

Abstract Mitochondrial DNA (mtDNA) diseases are multi‐systemic disorders caused by mutations affecting a fraction or the entirety of mtDNA copies. Currently, there no approved therapies for majority diseases. Challenges associated with engineering have in fact hindered study defects. Despite these difficulties, it has been possible to develop valuable cellular and animal models Here, we describe recent advances base editing generation three‐dimensional organoids from patient‐derived human‐induced pluripotent stem cells (iPSCs). Together already available modeling tools, combination novel technologies could allow determining impact specific distinct human cell types might help uncover how mutation load segregates during tissue organization. iPSC‐derived also represent platform identification treatment strategies probing vitro effectiveness gene therapies. These studies potential increase our mechanistic understanding may open way highly needed personalized therapeutic interventions.

Язык: Английский

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Large‐Area Field Potential Imaging Having Single Neuron Resolution Using 236 880 Electrodes CMOS‐MEA Technology

Advanced Science, Год журнала: 2023, Номер 10(20)

Опубликована: Апрель 23, 2023

Abstract The electrophysiological technology having a high spatiotemporal resolution at the single‐cell level and noninvasive measurements of large areas provide insights on underlying neuronal function. Here, complementary metal‐oxide semiconductor (CMOS)‐microelectrode array (MEA) is used that uses 236 880 electrodes each with an electrode size 11.22 × µm covering wide area 5.5 5.9 mm in presenting detailed single‐cell‐level neural activity analysis platform for brain slices, human iPS cell‐derived cortical networks, peripheral neurons, organoids. Propagation pattern characteristics between regions changes synaptic propagation into compounds based time‐series patterns, classification single DRG neuron firing patterns compound responses, axonal conduction to anticancer drugs, network activities transition organoids are extracted. This using CMOS‐MEA provides new understanding basic mechanisms circuits vitro ex vivo, neurological diseases drug discovery, toxicity assessment.

Язык: Английский

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Neural and metabolic dysregulation in PMM2-deficient human in vitro neural models

Cell Reports, Год журнала: 2024, Номер 43(3), С. 113883 - 113883

Опубликована: Март 1, 2024

Phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG) is a rare inborn error metabolism caused by deficiency the PMM2 enzyme, which leads to impaired protein glycosylation. While presents with primarily neurological symptoms, there limited knowledge about specific brain-related changes deficiency. Here, we demonstrate aberrant neural activity in 2D neuronal networks from PMM2-CDG individuals. Utilizing multi-omics datasets 3D human cortical organoids (hCOs) derived individuals, identify widespread decreases glycosylation, highlighting as key pathological feature PMM2-CDG, well mitochondrial structure and abnormal glucose PMM2-deficient hCOs, indicating disturbances energy metabolism. Correlation between enzymatic hCOs symptom severity suggests that level enzyme function directly influences manifestations. These findings enhance our understanding perturbations associated offering insights into underlying mechanisms potential directions for therapeutic interventions.

Язык: Английский

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The AMPK-related kinase NUAK1 controls cortical axons branching by locally modulating mitochondrial metabolic functions

Nature Communications, Год журнала: 2024, Номер 15(1)

Опубликована: Март 21, 2024

Abstract The cellular mechanisms underlying axonal morphogenesis are essential to the formation of functional neuronal networks. We previously identified autism-linked kinase NUAK1 as a central regulator axon branching through control mitochondria trafficking. However, (1) relationship between mitochondrial position, function and (2) downstream effectors whereby regulates remain unknown. Here, we report that recruitment synaptic boutons supports collateral branches stabilization rather than in mouse cortical neurons. deficiency significantly impairs metabolism ATP concentration, upregulation is sufficient rescue null neurons vitro vivo. Finally, found mitochondria-targeted microprotein BRAWNIN. Our results demonstrate exerts dual during its ability distribution metabolic activity.

Язык: Английский

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