Ramani Anand, Pasquini Giovanni, Gerkau Niklas J, Jadhav Vaibhav, Vinchure Omkar Suhas, Altinisik Nazlican, Windoffer Hannes, Muller Sarah, Rothenaigner Ina, Lin Sean, Mariappan Aruljothi, Rathinam Dhanasekaran, Mirsaidi Ali, Goureau Olivier, Ricci-Vitiani Lucia, D'Alessandris Quintino Giorgio, Wollnik Bernd, Muotri Alysson, Freifeld Limor, Jurisch-Yaksi Nathalie, Pallini Roberto, Rose Christine R, Busskamp Volker, Gabriel Elke, Hadian Kamyar, Gopalakrishnan Jay
Institute of Human Genetics, University Hospital, Friedrich-Schiller-Universität Jena, 07740, Jena, Germany.
Department of Ophthalmology, University Hospital Bonn, Medical Faculty, Bonn, Germany.
Nat Commun. 2024 Dec 19;15(1):10703. doi: 10.1038/s41467-024-55226-6.
Brain organoids offer unprecedented insights into brain development and disease modeling and hold promise for drug screening. Significant hindrances, however, are morphological and cellular heterogeneity, inter-organoid size differences, cellular stress, and poor reproducibility. Here, we describe a method that reproducibly generates thousands of organoids across multiple hiPSC lines. These High Quantity brain organoids (Hi-Q brain organoids) exhibit reproducible cytoarchitecture, cell diversity, and functionality, are free from ectopically active cellular stress pathways, and allow cryopreservation and re-culturing. Patient-derived Hi-Q brain organoids recapitulate distinct forms of developmental defects: primary microcephaly due to a mutation in CDK5RAP2 and progeria-associated defects of Cockayne syndrome. Hi-Q brain organoids displayed a reproducible invasion pattern for a given patient-derived glioma cell line. This enabled a medium-throughput drug screen to identify Selumetinib and Fulvestrant, as inhibitors of glioma invasion in vivo. Thus, the Hi-Q approach can easily be adapted to reliably harness brain organoids' application for personalized neurogenetic disease modeling and drug discovery.
脑类器官为大脑发育和疾病建模提供了前所未有的见解，并有望用于药物筛选。然而，显著的障碍包括形态和细胞异质性、类器官间大小差异、细胞应激以及可重复性差。在此，我们描述了一种可在多个诱导多能干细胞系中可重复地生成数千个类器官的方法。这些高数量脑类器官（Hi-Q脑类器官）展现出可重复的细胞结构、细胞多样性和功能，不存在异位激活的细胞应激途径，并允许冷冻保存和重新培养。患者来源的Hi-Q脑类器官重现了不同形式的发育缺陷：由CDK5RAP2突变导致的原发性小头畸形以及与早衰相关的科凯恩综合征缺陷。对于给定的患者来源的胶质瘤细胞系，Hi-Q脑类器官呈现出可重复的侵袭模式。这使得能够进行中通量药物筛选，以鉴定司美替尼和氟维司群作为体内胶质瘤侵袭的抑制剂。因此，Hi-Q方法可以很容易地进行调整，以可靠地利用脑类器官在个性化神经遗传疾病建模和药物发现中的应用。
