CircFGFR2 induces astrocyte pyroptosis to promote ischemic stroke.

Stroke is the second leading cause of death worldwide. Organoids, as disease models that closely mimic human physiology and pathology, are highly suitable for investigating the role of neural cells in brain diseases. However, there are few reports on circRNA research based on cerebral organoid stroke models. In this study, we established a human cerebral organoid ischemic stroke model induced by iPSCs and performed transcriptome sequencing. We found that at Days 31 and 52 of organoid development, the microglial marker Iba1, neuronal marker NeuN, and astrocytic marker GFAP exhibited strong fluorescent signals and showed co-localization with the proliferation marker KI67. The organoids showed high levels of the vascular marker CD31 and α-SMA, with co-localization between them. Transcriptome sequencing identified a total of 18 differentially expressed circRNAs (DEcircRNAs) and 2,111 differentially expressed genes (DEGs) in the human cerebral organoid stroke model. KEGG pathway enrichment analysis and WGCNA showed that these DEcircRNAs were involved in the pyroptosis pathway. Furthermore, we identified a pyroptosis-associated circRNA circFGFR2 who was up-regulated in astrocytes and promoted astrocyte pyroptosis. In summary, this study identified a highly expressed pyroptosis-associated circFGFR2 in the human cerebral organoid ischemic stroke model and demonstrated that circFGFR2 promoted astrocyte pyroptosis. This study provides new insights into the pathogenic mechanisms of ischemic stroke.