Ischemic stroke is a complex and heterogeneous neurological condition that represents a major cause of mortality and long-term disability worldwide. In addition to its functional consequences, stroke is frequently associated with cognitive impairment, further contributing to the clinical and socioeconomic burden of the disease [1]. Brain-derived neurotrophic factor (BDNF) is a neurotrophin that plays an important role in promoting neuronal survival, neuroprotection, and neuroplasticity [2]. Brain organoids are three-dimensional cellular model that recapitulate the cellular architecture and composition of the human brain and they represent a valuable platform to study ischemic stroke. In this work, we used a multi-omics framework to dissect the molecular effects of BDNF treatment in a cerebral organoid-based model of ischemic stroke. This integrated strategy allows the identification of condition-specific molecular signatures and pathways underlying ischemic injury and BDNF-mediated mechanisms [3]. Cerebral organoids were generated from human induced pluripotent stem cells (hiPSCs) and characterized over time using specific maturation markers. To establish an ischemic stroke model, organoids were exposed to different durations of oxygen-glucose deprivation (OGD) followed by reperfusion (R), allowing us to identify a sublethal condition suitable for evaluating the effects of BDNF treatment. We further characterized the organoid-based stroke model through Western Blotting, immunofluorescence, dPCR, and electrophysiological analyses, allowing an integrated assessment of cellular responses to ischemic injury. Furthermore, to investigate BDNF-modulated molecular mechanisms, transcriptomic profiling was performed using the Ion Torrent platform, while protein-level changes were assessed by Orbitrap-based mass spectrometry proteomics. The integration of these data aims to identify molecular signatures and pathways associated with ischemic injury and BDNF treatment. Overall, this multi-omics study provides new insights into the effects of BDNF treatment in a human three-dimensional model of ischemic stroke. 1. Li XY, Kong XM, Yang CH, et al. Global, regional, and national burden of ischemic stroke, 1990-2021: an analysis of data from the global burden of disease study 2021. EClinicalMedicine. 2024;75:102758. 2. Chen A, Xiong L, Tong Y and Mao M: The neuroprotective roles of BDNF in hypoxic ischemic brain injury (Review). Biomed Rep 1: 167-176, 2013. 3. Li W, Shao C, Zhou H, et al. Multi-omics research strategies in ischemic stroke: A multidimensional perspective. Ageing Res Rev. 2022;81:101730. Acknowledgements: This research was funded by Dompé farmaceutici S.p.A, by Progetto n. 74 – Piattaforma tecnologica integrata per l’identificazione e lo sviluppo di neurotrofine per il trattamento di patologie neurosensoriali a carico degli organi di vista e udito e patologie del CNS, rare o ad elevato bisogno di cura insoddisfatto (PINNACOLO) - CUP B19J23000180005 and by European Union-NextGenerationEU under the Italian University and Research (MUR) National Innovation Ecosystem grant ECS00000041-VITALITY-CUP E13C22001060006.
Exploring BDNF-mediated mechanisms in ischemic cerebral organoids through integrated multi-omics analysis
Fabrizio Ammannito;Chiara Giorgi;Michele d’Angelo;Annamaria Cimini
2026-01-01
Abstract
Ischemic stroke is a complex and heterogeneous neurological condition that represents a major cause of mortality and long-term disability worldwide. In addition to its functional consequences, stroke is frequently associated with cognitive impairment, further contributing to the clinical and socioeconomic burden of the disease [1]. Brain-derived neurotrophic factor (BDNF) is a neurotrophin that plays an important role in promoting neuronal survival, neuroprotection, and neuroplasticity [2]. Brain organoids are three-dimensional cellular model that recapitulate the cellular architecture and composition of the human brain and they represent a valuable platform to study ischemic stroke. In this work, we used a multi-omics framework to dissect the molecular effects of BDNF treatment in a cerebral organoid-based model of ischemic stroke. This integrated strategy allows the identification of condition-specific molecular signatures and pathways underlying ischemic injury and BDNF-mediated mechanisms [3]. Cerebral organoids were generated from human induced pluripotent stem cells (hiPSCs) and characterized over time using specific maturation markers. To establish an ischemic stroke model, organoids were exposed to different durations of oxygen-glucose deprivation (OGD) followed by reperfusion (R), allowing us to identify a sublethal condition suitable for evaluating the effects of BDNF treatment. We further characterized the organoid-based stroke model through Western Blotting, immunofluorescence, dPCR, and electrophysiological analyses, allowing an integrated assessment of cellular responses to ischemic injury. Furthermore, to investigate BDNF-modulated molecular mechanisms, transcriptomic profiling was performed using the Ion Torrent platform, while protein-level changes were assessed by Orbitrap-based mass spectrometry proteomics. The integration of these data aims to identify molecular signatures and pathways associated with ischemic injury and BDNF treatment. Overall, this multi-omics study provides new insights into the effects of BDNF treatment in a human three-dimensional model of ischemic stroke. 1. Li XY, Kong XM, Yang CH, et al. Global, regional, and national burden of ischemic stroke, 1990-2021: an analysis of data from the global burden of disease study 2021. EClinicalMedicine. 2024;75:102758. 2. Chen A, Xiong L, Tong Y and Mao M: The neuroprotective roles of BDNF in hypoxic ischemic brain injury (Review). Biomed Rep 1: 167-176, 2013. 3. Li W, Shao C, Zhou H, et al. Multi-omics research strategies in ischemic stroke: A multidimensional perspective. Ageing Res Rev. 2022;81:101730. Acknowledgements: This research was funded by Dompé farmaceutici S.p.A, by Progetto n. 74 – Piattaforma tecnologica integrata per l’identificazione e lo sviluppo di neurotrofine per il trattamento di patologie neurosensoriali a carico degli organi di vista e udito e patologie del CNS, rare o ad elevato bisogno di cura insoddisfatto (PINNACOLO) - CUP B19J23000180005 and by European Union-NextGenerationEU under the Italian University and Research (MUR) National Innovation Ecosystem grant ECS00000041-VITALITY-CUP E13C22001060006.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
