Deciphering Human Glioblastoma Invasion Using a Developmental Mature Rat Brain Organoid Model

Abstract

To model neurological diseases using brain organoids (BOs), there is a need for the development of terminally differentiated structures that reflect the structural and cellular complexity of mature brain tissue. Here we describe the developmental path of rat brain organoids (rBOs) into terminally differentiated brain structures. These organoids were compared to BOs derived from induced human pluripotent stem cells (iPSC) at the transcriptomic, proteomic and metabolomic level, showing that the rBOs present a higher degree of brain maturation. We further show that the rBOs can be used as an  ex vivo  avatar co-culture system to decipher important glioblastoma invasion parameters such as invasive cellular heterogeneity between patient-derived glioblastoma cells where key invasion parameters such as speed of single cell invasion and replacement of brain tissue by tumor cells can be measured and quantified in real-time. Finally, we show how this model can be used to assess therapeutic interventions, among others, directed toward neuro-gliomal α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor communication.

Funding Information: This work was supported by the Research Council of Norway, (Projects: 287033, 283790) to R. Bjerkvig the Norwegian Cancer Society (Projects: 220639, 208278) to R Bjerkvig, and the Luxembourg Research Fund (FNR) (PRIDE15/10675146 to SP Niclou). Wenjing Zhou, Xingang Li, and Jian Wang were supported by the Natural Science Foundation of China (81972351), the national “111” Project (B20058), and the Special Foundation for Taishan Scholars (tshw201502056 and ts20110814). CRUK core funding to the CRUK Beatson Institute (A31287) and CRUK funding to the Oncometabolism lab (A23982) to S. Tardito. We would like to thank Xiangyi Dong from the LCSB Metabolomics Platform, Luxembourg, for providing technical and analytical support. JM is supported by an FNR-ATTRACT grant (A18/BM/11809970).

Declaration of Interests: All authors declare that they have no known personal relationships or competing financial interests that affect the work reported in this paper.

Ethics Approval Statement: Ethical approval for establishing rBOs (approval number 19460 and for the establishment of orthotopic xenografts (approval number 18611) was given by the national Norwegian Food and Safety Authority. Euthanization was performed under isoflurane followed by cervical dislocation. Efforts were made to minimize animal suffering. Patient material was obtained from surgeries performed at the Haukeland University Hospital (Bergen, Norway). Written patient consent was obtained related to approved experimental procedures (project numbers 013.09, 2020/65185 and 151825; the Regional Ethics Committee, Bergen, Norway). All experiments in vivo were carried out in accordance with the Institutional Animal Care and Committee/Laboratory Animal Research Center protocols of the University of Bergen (UIB) following the ARRIVE guidelines.

Keywords: Brain organoids, glioblastoma, invasion, inhibition