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

Abstract

<jats:title>Abstract</jats:title><jats:p>Glioblastoma multiforme (GBM) are devastating neoplasms with high invasive capacity. GBM has been difficult to study <jats:italic>in vitro</jats:italic>. 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 <jats:italic>in vivo</jats:italic> 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 <jats:italic>in silico</jats:italic> 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 <jats:italic>in vitro</jats:italic>, with applications for both pharmacological screens and patient-specific treatment selection at a time scale amenable to clinical practice.</jats:p>

Cell Calcium Imaging as a Reliable Method to Study Neuron–Glial Circuits

“…Human organoids are also being developed to model glioblastoma, so invasion can be studied and how tumorigenesis relates to the extracellular matrix can be understood ( Silvia and Dai, 2020 ). Tumor cells within organoids extend a network of long microtubes, recapitulating the in vivo behavior of GBM ( Krieger et al, 2020 ). The transcriptional changes implicated in the invasion process are coherent across patient samples, indicating that GBM cells reactively upregulate genes required for their dispersion.…”

Organotypic Models to Study Human Glioblastoma: Studying the Beast in Its Ecosystem

“…Microfluidic organotypic cultures will not be included in this review. To the best of our knowledge, only six articles have been published on this topic (Nayernia et al, 2013;Ogawa et al, 2018;Bian et al, 2018aBian et al, , 2018bPlummer et al, 2019;Linkous et al, 2019;Krieger et al, 2020). cell culture approach went largely unnoticed and unrecognized-until this publication, it received only 15 citations.…”

“…The majority of human stem cell organotypic cultures used to generate GB-3DBrain models are the socalled cerebral organoids (Lancaster et al, 2013), either by using the exact protocol (Bian et al, 2018a;Ogawa et al, 2018) or by adding minor adaptations (Krieger et al, 2020;Linkous et al, 2019). The exception were Nayemia and collaborators, who used a different type of organotypic culture (the neural organoid) (Nayernia et al, 2013), and Plummer and collaborators, who utilized a model called BrainSpheres (Plummer et al, 2019) (Figure 1).…”

“…GB cells (directly from biopsies or patient-derived glioma cells) are added into the 3D structures. This primarily utilizes gravity approaches (Linkous et al, 2019;Krieger et al, 2020;Nayernia et al, 2013;Ogawa et al, 2018), allowing GB to attach cells to the surface of the organoid (Table 1). This could initially be viewed as an unfavorable approach, because contact between the GB with microenvironment cells will not be ideal.…”

“…This could initially be viewed as an unfavorable approach, because contact between the GB with microenvironment cells will not be ideal. However, Krieger and collaborators, by using a combination of live stained (or fixed) cerebral organoids embedded in Matrigel, together with clearing tissue technique and confocal imaging, were able to show tumor invasion after only 2 days of culture (Krieger et al, 2020). Tumoral cells added to the surface of the cerebral organoids were able to invade deeply into the tissue, exceeding 100 mm in the majority of organoids, with some cells detected at 300 mm from the organoid surface.…”