RESUMEN
Brain organoids represent a powerful tool for studying human neurological diseases, particularly those that affect brain growth and structure. However, many diseases manifest with clear evidence of physiological and network abnormality in the absence of anatomical changes, raising the question of whether organoids possess sufficient neural network complexity to model these conditions. Here, we explore the network-level functions of brain organoids using calcium sensor imaging and extracellular recording approaches that together reveal the existence of complex network dynamics reminiscent of intact brain preparations. We demonstrate highly abnormal and epileptiform-like activity in organoids derived from induced pluripotent stem cells from individuals with Rett syndrome, accompanied by transcriptomic differences revealed by single-cell analyses. We also rescue key physiological activities with an unconventional neuroregulatory drug, pifithrin-α. Together, these findings provide an essential foundation for the utilization of brain organoids to study intact and disordered human brain network formation and illustrate their utility in therapeutic discovery.
Asunto(s)
Encéfalo/fisiopatología , Epilepsia/fisiopatología , Neuronas , Adulto , Benzotiazoles/farmacología , Encéfalo/crecimiento & desarrollo , Señalización del Calcio , Preescolar , Epilepsia/diagnóstico por imagen , Femenino , Humanos , Células Madre Pluripotentes Inducidas , Proteína 2 de Unión a Metil-CpG/genética , Red Nerviosa/fisiopatología , Neurogénesis/genética , Neuroimagen , Síndrome de Rett/diagnóstico por imagen , Síndrome de Rett/fisiopatología , Análisis de la Célula Individual , Sinapsis , Tolueno/análogos & derivados , Tolueno/farmacología , TranscriptomaRESUMEN
The human cerebral cortex possesses distinct structural and functional features that are not found in the lower species traditionally used to model brain development and disease. Accordingly, considerable attention has been placed on the development of methods to direct pluripotent stem cells to form human brain-like structures termed organoids. However, many organoid differentiation protocols are inefficient and display marked variability in their ability to recapitulate the three-dimensional architecture and course of neurogenesis in the developing human brain. Here, we describe optimized organoid culture methods that efficiently and reliably produce cortical and basal ganglia structures similar to those in the human fetal brain in vivo. Neurons within the organoids are functional and exhibit network-like activities. We further demonstrate the utility of this organoid system for modeling the teratogenic effects of Zika virus on the developing brain and identifying more susceptibility receptors and therapeutic compounds that can mitigate its destructive actions.