Luminescent Human iPSC-Derived Neurospheroids Enable Modeling of Neurotoxicity After Oxygen–glucose Deprivation

Breedam, Elise Van; Nijak, Aleksandra; Buyle-Huybrecht, Tamariche; Stefano, Julia Di; Boeren, Marlies; Govaerts, Jonas; Quarta, Alessandra; Swartenbroekx, Tine; Jacobs, Eva; Menten, Björn; Gijsbers, Rik; Delputte, Peter; Alaerts, Maaike; Hassannia, Behrouz; Loeys, Bart; Berneman, Zwi; Timmermans, Jean‐Pierre; Berghe, Tom Vanden; Fransen, Erik; Wouters, An; Vos, Winnok H. De; Ponsaerts, Peter

doi:10.1007/s13311-022-01212-z

Cited by 12 publications

(10 citation statements)

References 47 publications

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“…More specifically, the response to treatment with the pan-caspase inhibitor Z-VAD-FMK during and after OGD differed between NSCs cultured in 2D versus NSC-derived neurospheroids. Where Z-VAD-FMK conferred neuroprotection in 2D, in line with other publications, it failed to protect neurospheroids under OGD [149]. Altogether, these findings further underscore the importance of 3D models in basic as well as applied in vitro stroke research to complement conventional 2D cell cultures and in vivo animal studies.…”

Section: Dimensionality Of Cell Culture Models For In Vitro Ischaemic...supporting

confidence: 85%

“…The results of the last-mentioned study were also replicated by our own studies. With the aim of developing a human neurospheroid model for ischaemic stroke, we equipped iPSC-derived neurospheroids with intrinsic bioluminescence to enable the real-time monitoring of the viability of neurospheroids subjected to OGD and were able to model OGD-mediated neurotoxicity [149]. By comparing 1-week-old with 4-week-old neurospheroids, containing a high proportion of undifferentiated NSCs and intermediate progenitors/immature neurons, respectively, it was demonstrated that 1-week-old neurospheroids were able to completely and spontaneously recover from the initial OGD-induced damage over the course of one week, unlike 4-week-old neurospheroids.…”

Section: Dimensionality Of Cell Culture Models For In Vitro Ischaemic...mentioning

confidence: 99%

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Promising Strategies for the Development of Advanced In Vitro Models with High Predictive Power in Ischaemic Stroke Research

Breedam

Ponsaerts

2022

IJMS

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Although stroke is one of the world’s leading causes of death and disability, and more than a thousand candidate neuroprotective drugs have been proposed based on extensive in vitro and animal-based research, an effective neuroprotective/restorative therapy for ischaemic stroke patients is still missing. In particular, the high attrition rate of neuroprotective compounds in clinical studies should make us question the ability of in vitro models currently used for ischaemic stroke research to recapitulate human ischaemic responses with sufficient fidelity. The ischaemic stroke field would greatly benefit from the implementation of more complex in vitro models with improved physiological relevance, next to traditional in vitro and in vivo models in preclinical studies, to more accurately predict clinical outcomes. In this review, we discuss current in vitro models used in ischaemic stroke research and describe the main factors determining the predictive value of in vitro models for modelling human ischaemic stroke. In light of this, human-based 3D models consisting of multiple cell types, either with or without the use of microfluidics technology, may better recapitulate human ischaemic responses and possess the potential to bridge the translational gap between animal-based in vitro and in vivo models, and human patients in clinical trials.

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Section: Dimensionality Of Cell Culture Models For In Vitro Ischaemic...supporting

confidence: 85%

Section: Dimensionality Of Cell Culture Models For In Vitro Ischaemic...mentioning

confidence: 99%

Promising Strategies for the Development of Advanced In Vitro Models with High Predictive Power in Ischaemic Stroke Research

Breedam

Ponsaerts

2022

IJMS

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“…30,31 The vulnerability of differentiating cells to OGD was also confirmed recently by Van Breedam and colleagues, who developed a luminescent human iPSC-derived neurospheroid model that allows real-time measurement of neural viability under OGD. 32 By this approach the authors demonstrated that OGD-mediated injury was reversible in younger (1-week-old), but not in more mature (4-week-old) neurospheroids, suggesting the need for longer neurospheroid maturation for in vitro stroke research. Pas¸ca et al identified impairments in neuron progenitor distribution in specific areas of brain spheroids cultured in hypoxic conditions, 11 but no toxic effects on differentiated neurons were reported.…”

Section: Discussionmentioning

confidence: 98%

Neural cortical organoids from self-assembling human iPSC as a model to investigate neurotoxicity in brain ischemia

Paola

Pischiutta

Comolli

et al. 2023

J Cereb Blood Flow Metab

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Brain ischemia is a common acute injury resulting from impaired blood flow to the brain. Translation of effective drug candidates from experimental models to patients has systematically failed. The use of human induced pluripotent stem cells (iPSC) offers new opportunities to gain translational insights into diseases including brain ischemia. We used a human 3D self-assembling iPSC-derived model (human cortical organoids, hCO) to characterize the effects of ischemia caused by oxygen-glucose deprivation (OGD). hCO exposed to 2 h or 8 h of OGD had neuronal death and impaired neuronal network complexity, measured in whole-mounting microtubule-associated protein 2 (MAP-2) immunostaining. Neuronal vulnerability was reflected by a reduction in MAP-2 mRNA levels, and increased release of neurofilament light chain (NfL) in culture media, proportional to OGD severity. Glial fibrillary acidic protein (GFAP) gene or protein levels did not change in hCO, but their release in medium increased after prolonged OGD. In conclusion, this human 3D iPSC-based in vitro model of brain ischemic injury is characterized by marked neuronal injury reflected by the release of the translational biomarker NfL which is relevant for testing neuroprotective strategies.

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“…Neurospheroids were formed as described previously [26]. Briefly, eGFP/Luc human iPSC-NSCs were seeded at equal densities of 1.6 x 10 4 cells per well (ULA 96-well plate (Corning)) in neural expansion medium (NEM, Gibco).…”

Section: Methodsmentioning

confidence: 99%

“…After fixation, neurospheroids were processed as described previously to allow high-throughput staining [26]. Briefly, a silicone mold with 66 wells corresponding to the size of the neurospheroids was filled with Tissue-Tek-OCT (VWR).…”

Section: Methodsmentioning

confidence: 99%

Amelioration of Functional and Histopathological Consequences after Spinal Cord Injury through Phosphodiesterase 4D (PDE4D) Inhibition

Schepers,

Hendrix,

Mussen

et al. 2023

Preprint

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Spinal cord injury (SCI) severely impacts the patient's quality of life. Two key strategies are currently being considered to improve clinical outcomes after SCI: modulation of the neuroinflammatory response, which exacerbates the primary injury, and stimulation of neuro-regenerative repair mechanisms to improve functional recovery. Cyclic adenosine monophosphate (cAMP) is a second messenger involved in both processes. Following SCI, intracellular levels of cAMP are known to decrease over time. Therefore, preventing cAMP degradation represents a promising strategy to suppress inflammation while stimulating regeneration. Intracellular cAMP levels are controlled by its hydrolyzing enzymes phosphodiesterases (PDEs). The PDE4 family is most abundantly expressed in the central nervous system (CNS) and its inhibition has been shown to be therapeutically relevant for managing SCI pathology. Unfortunately, the use of full PDE4 inhibitors at therapeutic doses is associated with severe emetic side effects, hampering their translation toward clinical applications. Therefore, in this study, we evaluated the effect of inhibiting specific PDE4 subtypes (PDE4B and PDE4D) on inflammatory and regenerative processes following SCI, as inhibitors selective for these subtypes have been demonstrated to be well-tolerated. We reveal that administration of the PDE4D inhibitor Gebr32a, but not the PDE4B inhibitor A33, improved functional as well as histopathological outcomes after SCI, comparable to results obtained with the full PDE4 inhibitor roflumilast. Furthermore, using a luminescent human iPSC-derived neurospheroid model, we show that PDE4D inhibition stabilizes neural viability by preventing apoptosis and stimulating neuronal differentiation. These findings strongly suggest that specific PDE4D inhibition offers a novel therapeutic approach for SCI.

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Luminescent Human iPSC-Derived Neurospheroids Enable Modeling of Neurotoxicity After Oxygen–glucose Deprivation

Cited by 12 publications

References 47 publications

Promising Strategies for the Development of Advanced In Vitro Models with High Predictive Power in Ischaemic Stroke Research

Promising Strategies for the Development of Advanced In Vitro Models with High Predictive Power in Ischaemic Stroke Research

Neural cortical organoids from self-assembling human iPSC as a model to investigate neurotoxicity in brain ischemia

Amelioration of Functional and Histopathological Consequences after Spinal Cord Injury through Phosphodiesterase 4D (PDE4D) Inhibition

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