Glioblastoma Model Using Human Cerebral Organoids

Ogawa, Junko; Pao, Gerald M.; Shokhirev, Maxim N.; Verma, Inder M.

doi:10.1016/j.celrep.2018.03.105

Cited by 307 publications

(343 citation statements)

References 20 publications

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“…Previous in vitro models have suffered from limited physiological relevance or have been incompatible with the time scales for clinical decision-making 6 . Recent studies have shown that human cerebral organoids can be used as a platform for tumor cell transplantation or genetic engineering of tumors, enabling microscopic observation of tumor development [13][14][15] . However, tumor cell interactions with normal brain cells have not been addressed yet.…”

Section: Introductionmentioning

confidence: 99%

Modeling glioblastoma invasion using human brain organoids and single-cell transcriptomics

Krieger

Tirier

Park

et al. 2019

Preprint

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AbstractGlioblastoma multiforme (GBM) are devastating neoplasms with high invasive capacity. GBM has been difficult to study in vitro. Therapeutic progress is also limited by cellular heterogeneity within and between tumors. To address these challenges, we present an experimental model using human cerebral organoids as a scaffold for patient-derived glioblastoma cell invasion. By tissue clearing and confocal microscopy, we show that tumor cells within organoids extend a network of long microtubes, recapitulating the in vivo behavior of GBM. Single-cell RNA-seq of GBM cells before and after co-culture with organoid cells reveals transcriptional changes implicated in the invasion process that are coherent across patient samples, indicating that GBM cells reactively upregulate genes required for their dispersion. Functional therapeutic targets are identified by an in silico receptor-ligand pairing screen detecting potential interactions between GBM and organoid cells. Taken together, our model has proven useful for studying GBM invasion and transcriptional heterogeneity in vitro, with applications for both pharmacological screens and patient-specific treatment selection at a time scale amenable to clinical practice.

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Section: Introductionmentioning

confidence: 99%

Modeling glioblastoma invasion using human brain organoids and single-cell transcriptomics

Krieger

Tirier

Park

et al. 2019

Preprint

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“…However, PDX models are restricted by variable engraftment efficiency, mixing with animal cells, limited throughput, and long latency in tumor generation, which limits their usefulness for testing personalized therapies 27 . Cerebral organoids, self-organizing 3D cultures that resemble the developing brain 28 , have been genetically altered to develop oncogenic properties or co-cultured with tumor spheres to model tumor cell invasion 29,30 . While cerebral organoids are useful, readily manipulatable models, they represent only the fetal brain without important non-neural cell types, such as immune and endothelial cells, and do not recapitulate the cellular diversity or architecture of glioblastoma tumors.…”

Section: Development Of Glioblastoma Organoid Culture and Biobankingmentioning

confidence: 99%

Generation and biobanking of patient-derived glioblastoma organoids and their application in CAR-T cell testing

Jacob

Ming

Song

2020

Preprint

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Short Summary: Detailed procedures for generating and biobanking glioblastoma organoids from resected patient tumor tissue and testing CAR-T cell efficacy by co-culture. Additional procedures for tissue processing, immunohistology, and detecting hypoxia gradients and actively proliferating cells. AbstractGlioblastoma tumors exhibit extensive inter-and intra-tumoral heterogeneity, which has contributed to the poor outcomes of numerous clinical trials and continues to complicate the development of effective therapeutic strategies. Current in vitro models do not preserve the cellular and mutational diversity of parent tumors and often require a lengthy generation time with variable efficiency. Here, we describe detailed procedures for generating glioblastoma organoids (GBOs) from surgically resected patient tumor tissue using a chemically defined medium without cell dissociation. By preserving cell-cell interactions and minimizing clonal selection, GBOs maintain the cellular heterogeneity of parent tumors. We include methods for passaging and cryopreserving GBOs for continued use, biobanking, and long-term recovery. We further describe procedures for investigating patient-specific responses to immunotherapies by co-culturing GBOs with chimeric antigen receptor (CAR) T cells. This protocol takes approximately 2-4 weeks to generate GBOs and 5-7 days to perform CAR-T cell co-culture. Competence with human cell culture, tissue processing, and immunohistology is required for optimal results.An ideal in vitro model for glioblastoma would maintain cellular diversity, preserve native cell-cell interactions, recapitulate defining histological characteristics, and maintain the transcriptomes and mutation profiles of parent tumors. To be useful for personalized medicine approaches, the model would need a quick generation time with high fidelity for success and be scalable to test potential therapeutics. This would require maintaining patient-derived glioblastoma tissue in culture with minimal perturbation. Towards this goal, we opted to directly culture micro-dissected tumor pieces without cell dissociation to preserve cell-cell interactions and minimize clonal selection. We start from resected patient tumor tissue that can be sampled from different tumor foci or subdivided into regional samples to analyze intra-tumoral heterogeneity. Through testing of many medium formulations, we learned that tumor pieces did not require basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), serum, or added extracellular matrix such as Matrigel to survive, form organoids, and proliferate at rates similar to parent tumors 11 . We optimized a chemically defined medium that contains basic nutrients and components needed to support glial cell health with few exogenous factors to minimize clonal selection induced by growth factors and decrease potential treatment confounders. This resulted in an optimized version of our medium used to maintain human brain organoid cultures 12,13 . We culture tumor pieces on an orbital shaker to inc...

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“…While this study already proves the potential of brain organoids to help understand tumorigenesis, variations in expression levels of oncogenes from organoid to organoid requires further refinement of the methodology. Ogawa et al devised an elegant method to study the invasion behavior of patient‐derived glioblastoma cell lines in brain organoids and identified that GBM cells have the ability to invade within the 3D neuroepithelium. This method basically establishes human brain organoids as a platform to further characterize the invasive behavior of patient‐derived GBM cells.…”

Section: Expectations From the Current State‐of‐the‐art Organoidsmentioning

confidence: 99%

The Emergence of Stem Cell‐Based Brain Organoids: Trends and Challenges

Gopalakrishnan

2019

BioEssays

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Recent developments in 3D cultures exploiting the self‐organization ability of pluripotent stem cells have enabled the generation of powerful in vitro systems termed brain organoids. These 3D tissues recapitulate many aspects of human brain development and disorders occurring in vivo. When combined with improved differentiation methods, these in vitro systems allow the generation of more complex “assembloids,” which are able to reveal cell diversities, microcircuits, and cell–cell interactions within their 3D organization. Here, the ways in which human brain organoids have contributed to demystifying the complexities of brain development and modeling of developmental disorders is reviewed and discussed. Furthermore, challenging questions that are yet to be addressed by emerging brain organoid research are discussed.

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Glioblastoma Model Using Human Cerebral Organoids

Cited by 307 publications

References 20 publications

Modeling glioblastoma invasion using human brain organoids and single-cell transcriptomics

Modeling glioblastoma invasion using human brain organoids and single-cell transcriptomics

Generation and biobanking of patient-derived glioblastoma organoids and their application in CAR-T cell testing

The Emergence of Stem Cell‐Based Brain Organoids: Trends and Challenges

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