An in silico and in vitro human neuronal network model reveals cellular mechanisms beyond NaV1.1 underlying Dravet syndrome DOI Creative Commons
Nina Doorn, Eline van Hugte, Ummi Ciptasari

et al.

Stem Cell Reports, Journal Year: 2023, Volume and Issue: 18(8), P. 1686 - 1700

Published: July 6, 2023

Human induced pluripotent stem cell (hiPSC)-derived neuronal networks on multi-electrode arrays (MEAs) provide a unique phenotyping tool to study neurological disorders. However, it is difficult infer cellular mechanisms underlying these phenotypes. Computational modeling can utilize the rich dataset generated by MEAs, and advance understanding of disease mechanisms. existing models lack biophysical detail, or validation calibration relevant experimental data. We developed in silico model that accurately simulates healthy MEAs. To demonstrate potential our model, we studied derived from Dravet syndrome (DS) patient with missense mutation SCN1A, encoding sodium channel NaV1.1. Our revealed dysfunctions were insufficient replicate vitro DS phenotype, predicted decreased slow afterhyperpolarization synaptic strengths. verified changes patient-derived neurons, demonstrating utility predict

Language: Английский

Human neuronal networks on micro-electrode arrays are a highly robust tool to study disease-specific genotype-phenotype correlations in vitro DOI Creative Commons
Britt Mossink, Anouk H. A. Verboven, Eline van Hugte

et al.

Stem Cell Reports, Journal Year: 2021, Volume and Issue: 16(9), P. 2182 - 2196

Published: July 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.

Language: Английский

Citations

98
Imbalanced autophagy causes synaptic deficits in a human model for neurodevelopmental disorders DOI Creative Commons
Katrin Linda, Elly Lewerissa, Anouk H. A. Verboven

et al.

Autophagy, Journal Year: 2021, Volume and Issue: 18(2), P. 423 - 442

Published: July 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.

Language: Английский

Citations

58
iPSC models of mitochondrial diseases DOI Creative Commons

Sonja Heiduschka,

Alessandro Prigione

Neurobiology of Disease, Journal Year: 2025, Volume and Issue: unknown, P. 106822 - 106822

Published: Jan. 1, 2025

Language: Английский

Citations

1
A MYT1L syndrome mouse model recapitulates patient phenotypes and reveals altered brain development due to disrupted neuronal maturation DOI Creative Commons
Jiayang Chen,

Mary E. Lambo,

Xia Ge

et al.

Neuron, Journal Year: 2021, Volume and Issue: 109(23), P. 3775 - 3792.e14

Published: Oct. 5, 2021

Language: Английский

Citations

56
Pathogenic mitochondrial DNA 3243A>G mutation: From genetics to phenotype DOI Creative Commons
Danyang Li,

Chunmei Liang,

Tao Zhang

et al.

Frontiers in Genetics, Journal Year: 2022, Volume and Issue: 13

Published: Oct. 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.

Language: Английский

Citations

35
Modeling mitochondrial DNA diseases: from base editing to pluripotent stem‐cell‐derived organoids DOI Creative Commons
Isabella Tolle, Valeria Tiranti, Alessandro Prigione

et al.

EMBO Reports, Journal Year: 2023, Volume and Issue: 24(4)

Published: March 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.

Language: Английский

Citations

20
The potential of in vitro neuronal networks cultured on micro electrode arrays for biomedical research DOI Creative Commons

Marta Cerina,

Maria Carla Piastra, Monica Frega

et al.

Progress in Biomedical Engineering, Journal Year: 2023, Volume and Issue: 5(3), P. 032002 - 032002

Published: April 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.

Language: Английский

Citations

20
Large‐Area Field Potential Imaging Having Single Neuron Resolution Using 236 880 Electrodes CMOS‐MEA Technology DOI Creative Commons
Ikuro Suzuki, Naoki Matsuda,

Xiaobo Han

et al.

Advanced Science, Journal Year: 2023, Volume and Issue: 10(20)

Published: April 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.

Language: Английский

Citations

17
Neural and metabolic dysregulation in PMM2-deficient human in vitro neural models DOI Creative Commons
Silvia Radenkovic, Rohit Budhraja, Teun M. Klein Gunnewiek

et al.

Cell Reports, Journal Year: 2024, Volume and Issue: 43(3), P. 113883 - 113883

Published: March 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.

Language: Английский

Citations

8
The AMPK-related kinase NUAK1 controls cortical axons branching by locally modulating mitochondrial metabolic functions DOI Creative Commons
Marine Lanfranchi,

Sozerko Yandiev,

Géraldine Meyer-Dilhet

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: March 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.

Language: Английский

Citations

7