An in silico and in vitro human neuronal network model reveals cellular mechanisms beyond NaV1.1 underlying Dravet syndrome

Doorn, Nina; Hugte, Eline J.H. van; Ciptasari, Ummi; Mordelt, Annika; Meijer, Hil Gaétan Ellart; Schubert, Dirk W.; Frega, Monica; Kasri, Nael Nadif; Putten, Michel J.A.M. van

doi:10.1016/j.stemcr.2023.06.003

Cited by 5 publications

(23 citation statements)

References 64 publications

(96 reference statements)

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“…To narrow down the possible cellular mechanisms that underlie the observed altered network synchronization, we employed a recently developed predictive and hypothesis generating in silico model. 23 The cellular mechanisms identified by the in silico CACNA1A +/model were 1) decreased synaptic function with an increased NMDA/AMPA ratio, and 2) decreased sodium and potassium channel function. We tested and validated these findings in our in vitro CACNA1A +/model and indeed found a decrease in synaptic function with enhanced contribution of GluA2-lacking AMPA receptors, and increased intrinsic excitability through reduced potassium channel function.…”

Section: Discussionmentioning

confidence: 99%

“…Network bursts were detected using a firing rate threshold method as described earlier. 23 From this, the network burst frequency, network burst duration, and network burst percentage features were calculated. The correlation coefficients between the binary spike train signals of all the electrodes were calculated to define the average correlation and standard deviation of the correlation features.…”

Section: In Silico Modelingmentioning

confidence: 99%

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CACNA1Ahaploinsufficiency leads to reduced synaptic function and increased intrinsic excitability

Hommersom,

Doorn,

Puvogel

et al. 2024

Preprint

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Haploinsufficiency of theCACNA1Agene, encoding the pore-forming α1 subunit of P/Q-type voltage-gated calcium channels, is associated with a clinically variable phenotype ranging from cerebellar ataxia, to neurodevelopmental syndromes with epilepsy and intellectual disability.To understand the pathological mechanisms ofCACNA1Aloss-of-function variants, we characterized a human neuronal model forCACNA1Ahaploinsufficiency, by differentiating isogenic induced pluripotent stem cell lines into glutamatergic neurons, and investigated the effect ofCACNA1Ahaploinsufficiency on mature neuronal networks through a combination of electrophysiology, gene expression analysis, andin silicomodeling.We observed an altered network synchronization inCACNA1A+/−networks alongside synaptic deficits, notably marked by an augmented contribution of GluA2-lacking AMPA receptors. Intriguingly, these synaptic perturbations coexisted with increased non-synaptically driven activity, as characterized by inhibition of NMDA and AMPA receptors on micro-electrode arrays. Single-cell electrophysiology and gene expression analysis corroborated this increased intrinsic excitability through reduced potassium channel function and expression. Moreover, we observed partial mitigation of theCACNA1A+/−network phenotype by 4-aminopyridine, a therapeutic intervention for episodic ataxia type 2.In summary, our study pioneers the characterization of a human induced pluripotent stem cell-derived neuronal model forCACNA1Ahaploinsufficiency, and has unveiled novel mechanistic insights. Beyond showcasing synaptic deficits, this neuronal model exhibited increased intrinsic excitability mediated by diminished potassium channel function, underscoring its potential as a therapeutic discovery platform with predictive validity.

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Section: Discussionmentioning

confidence: 99%

Section: In Silico Modelingmentioning

confidence: 99%

CACNA1Ahaploinsufficiency leads to reduced synaptic function and increased intrinsic excitability

Hommersom,

Doorn,

Puvogel

et al. 2024

Preprint

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“…Next, we aimed to investigate the mechanisms underlying the appearance of fragmented NBs in patient-derived neuronal networks. For this, we employed our previously developed computational model of hiPSC-derived excitatory neuronal networks on MEA [7], which has been shown to successfully identify the effect of specific cellular changes on the network activity.…”

Section: Our Biophysical Models Replicate Fragmented Nbs With Increas...mentioning

confidence: 99%

“…The realistic in silico model is described in Doorn et al [7]. In short, it consists of one hundred HH-type neurons with voltage-gated sodium and potassium channels, as well as leaky channels and sAHP channels, modeled as a potassium channel whose conductance increases upon action potential (AP) firing.…”

Section: Realistic In Silico Modelmentioning

confidence: 99%

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Breaking the Burst: Unveiling Mechanisms Behind Fragmented Network Bursts in Patient-derived Neurons

Doorn,

Voogd,

Levers

et al. 2024

Preprint

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Fragmented network bursts (NBs) are observed as a phenotypic driver in many patient-derived neuronal networks on multi-electrode arrays (MEAs), but the pathophysiological mechanisms underlying this phenomenon are unknown. Here, we used our previously developed biophysically detailedin silicomodel to investigate these mechanisms. Fragmentation of NBs in our model simulations occurred only when the level of short-term synaptic depression (STD) was enhanced, suggesting that STD is a key player. Experimental validation with Dynasore, an STD enhancer, induced fragmented NBs in healthy neuronal networksin vitro. Additionally, we showed that strong asynchronous neurotransmitter release, NMDA currents, or short-term facilitation (STF) can support the emergence of multiple fragments in NBs by producing excitation that persists after high-frequency firing stops. Our results provide important insights into disease mechanisms and potential pharmaceutical targets for neurological disorders modeled using hiPSC-derived neurons.

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Breaking the burst: Unveiling mechanisms behind fragmented network bursts in patient-derived neurons

Doorn,

Voogd,

Levers

et al. 2024

Stem Cell Reports

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An in silico and in vitro human neuronal network model reveals cellular mechanisms beyond NaV1.1 underlying Dravet syndrome

Cited by 5 publications

References 64 publications

CACNA1Ahaploinsufficiency leads to reduced synaptic function and increased intrinsic excitability

CACNA1Ahaploinsufficiency leads to reduced synaptic function and increased intrinsic excitability

Breaking the Burst: Unveiling Mechanisms Behind Fragmented Network Bursts in Patient-derived Neurons

Breaking the burst: Unveiling mechanisms behind fragmented network bursts in patient-derived neurons

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