Humanized cerebral organoids-based ischemic stroke model for discovering of potential anti-stroke agents
Humanized cerebral organoids-based ischemic stroke model for discovering of potential anti-stroke agents
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DOI：
10.1038/s41401-022-00986-4
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发表时间：
2022-09
影响因子：
8.2
通讯作者：
Shu-Na Wang;Zhi Wang;Xi-Yuan Wang;Xiu-Ping Zhang;Tian-ying Xu;C. Miao
中科院分区：
文献类型：
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作者：
Shu-Na Wang;Zhi Wang;Xi-Yuan Wang;Xiu-Ping Zhang;Tian-ying Xu;C. Miao
建立更符合人脑生理和功能的斯托克实验模型是开发有效抗脑卒中药物的瓶颈。由人多能干细胞构建的三维脑类器官（COs）可以模拟在体人脑的细胞组成、皮质结构、脑神经连接和表观遗传基因组学，为建立人源化缺血性脑卒中模型提供了良好的应用前景。CO已被用于低氧条件下的缺氧损伤模型，但在体外氧糖剥夺（OGD）诱导的缺血性脑卒中损伤中CO的变化及其在抗脑卒中药物筛选中的应用尚未见报道。本研究比较了不同培养时间COs的细胞组成，并探讨了85 d时COs的细胞类型、细胞比例和体积大小。选用直径大于2 mm的85 d-CO建立人源化缺血性脑卒中OGD模型。通过测定模型的时间-损伤关系，观察到COs缺血性损伤随OGD暴露时间的增加而加重，为不同OGD条件下COs的损伤程度获得第一手证据。通过经证实的泛半胱天冬酶抑制剂Z-VAD-FMK（20 μM）和Bcl-2抑制剂navitoclax（0.5 μM）验证了模型对缺血性损伤和相关治疗的敏感性。神经保护剂依达拉奉、丁苯酞、P7 C3-A20和ZL 006（各10 μM）在该模型中发挥了相似的有益作用。综上所述，本研究建立了一种基于COs的人源化缺血性脑卒中模型，为抗脑卒中药物开发提供了新的研究平台。
Establishing a stoke experimental model, which is better in line with the physiology and function of human brain, is the bottleneck for the development of effective anti-stroke drugs. A three-dimensional cerebral organoids (COs) from human pluripotent stem cells can mimic cell composition, cortical structure, brain neural connectivity and epigenetic genomics of in-vivo human brain, which provides a promising application in establishing humanized ischemic stroke model. COs have been used for modeling low oxygen condition-induced hypoxic injury, but there is no report on the changes of COs in response to in vitro oxygen-glucose deprivation (OGD)-induced damage of ischemic stroke as well as its application in testing anti-stroke drugs. In this study we compared the cell composition of COs at different culture time and explored the cell types, cell ratios and volume size of COs at 85 days (85 d-CO). The 85 d-CO with diameter more than 2 mm was chosen for establishing humanized ischemic stroke model of OGD. By determining the time-injury relationship of the model, we observed aggravated ischemic injury of COs with OGD exposure time, obtaining first-hand evidence for the damage degree of COs under different OGD condition. The sensitivity of the model to ischemic injury and related treatment was validated by the proven pan-Caspase inhibitor Z-VAD-FMK (20 μM) and Bcl-2 inhibitor navitoclax (0.5 μM). Neuroprotective agents edaravone, butylphthalide, P7C3-A20 and ZL006 (10 μM for each) exerted similar beneficial effects in this model. Taken together, this study establishes a humanized ischemic stroke model based on COs, and provides evidence as a new research platform for anti-stroke drug development.