Biobanked Glioblastoma Patient-Derived Organoids as a Precision Medicine Model to Study Inhibition of Invasion

Abstract

Glioblastoma (GBM) is highly resistant to treatment and invasion into the surrounding brain is a cancer hallmark that leads to recurrence despite surgical resection. With the emergence of precision medicine, patient-derived 3D systems are considered potentially robust GBM preclinical models. In this study, we screened a library of 22 anti-invasive compounds (i.e., NF-kB, GSK-3-B, COX-2, and tubulin inhibitors) using glioblastoma U-251 MG cell spheroids. We evaluated toxicity and invasion inhibition using a 3D Matrigel invasion assay. We next selected three compounds that inhibited invasion and screened them in patient-derived glioblastoma organoids (GBOs). We developed a platform using available macros for FIJI/ImageJ to quantify invasion from the outer margin of organoids. Our data demonstrated that a high-throughput invasion screening can be done using both an established cell line and patient-derived 3D model systems. Tubulin inhibitor compounds had the best efficacy with U-251 MG cells, however, in ex vivo patient organoids the results were highly variable. Our results indicate that the efficacy of compounds is highly related to patient intra and inter-tumor heterogeneity. These results indicate that such models can be used to evaluate personal oncology therapeutic strategies.

Metabolic-imaging of human glioblastoma live tumors: A new precision-medicine approach to predict tumor treatment response early

“…Recent advances in organoid technology have revolutionized in vitro culture tools for biomedical research by creating powerful 3D-dimensional models, that better preserve the local cytoarchitecture and native cell-cell interactions of original tumors (5). Despite numerous ex-vivo drug testing approaches leverage on 3d-in vitro GB models, they still have shortcomings and none fully captures the complexity of each individual glioblastoma cellular organization and composition overlooking therefore the relevance of how the tumor microenvironment affects tumor behavior and drug response (6)(7)(8). These drug testing approaches have several limiting requirements such as: a) extended time of performance that leads to a long period of in vitro tumor culturing with a consequent molecular and morphological transformation diverging from the parental tumor in vivo (6); b) technical measurements requiring dissociation of the original tumoral tissue down to a single cell suspension, losing therefore the tumor cytoarchitecture and cell-cell interaction characteristics (8); c) use of non-human animal models which implies laborsome procedures and introduction of biases due to host organism-tumor interactions (9).…”

“…Here in this study, to answer these requirements, we offer a unique novel treatment-testing approach in patient-derived glioblastoma 3D organoids. We first developed a protocol to generate an in vitro vital patient-derived IDH1/2 wild-type GB 3D model that we termed "glioblastoma explant" (GB-EXP), which, unlike other models (6)(7)(8), is minimally handled, briefly grown in culture with no animal involvement, no dissociation and passaging to preserve the parental cytoarchitecture. To build a treatment-response predictor tool, we applied an imaging method called FLIM that exploits the intrinsic auto-fluorescence molecular properties of NAD(P)H, a metabolic enzymatic cofactor, that is associated with the metabolic state of the cell/ tissue.…”

Receptor tyrosine kinase (RTK) targeting in pediatric high-grade glioma and diffuse midline glioma: Pre-clinical models and precision medicine

“…GBOs have been shown to have similar histological and transcriptional profiles to their respective patient tumor and recapitulate patient-specific responses and resistance to standard of care therapy ( 108 , 109 ). A cryopreserved GBO repository has been developed and used to screen a library of 22 compounds that inhibit tumor invasion into surrounding tissue ( 110 ). GBOs have also been used to study response to chimeric antigen T (CAR-T) cell immunotherapy in the setting of the EGFRvIII variant ( 111 ), mTOR inhibitors in PTEN loss ( 112 ), and STAT inhibitors in Li-Fraumeni syndrome ( 113 ).…”

Metabolic-imaging of human glioblastoma live tumors: a new precision-medicine approach to predict tumor treatment response early

“…To overcome these limitations, we first developed a protocol to generate an in-vitro vital patient-derived IDH1/2 wild-type GB 3D model that we termed “glioblastoma explant” (GB-EXP), which, unlike other models 5 6 , is minimally handled and shortly grown in culture with no passaging in order to preserve the parental cyto-architecture. Then, to build a drug-response predictor tool, we combined NADH fluorescence lifetime imaging microscopy (FLIM) and the GB-EXP model to detect metabolic changes that are increasingly used to predict drug efficacy and to detect early responses to cancer treatment 7 8 9 .…”

“…Recent advances in organoid technology have revolutionized in vitro culture tools for biomedical research by creating powerful 3D-dimensional models, that better preserve the local cytoarchitecture and native cell-cell interactions of original tumors 5 . Despite numerous ex-vivo drug testing approaches leverage on 3d-in vitro GB models, they still have shortcomings and none fully captures the complexity of each individual glioblastoma cellular organization and composition overlooking therefore the relevance of how the tumor microenvironment affects tumor behavior and drug response 6 7 8 . These drug testing approaches have several limiting requirements such as: a) extended time of performance that leads to a long period of in vitro tumor culturing with a consequent molecular and morphological transformation diverging from the parental tumor in vivo 6 ; b) technical measurements requiring dissociation of the original tumoral tissue down to a single cell suspension, losing therefore the tumor cytoarchitecture and cell-cell interaction characteristics 8 ; c) use of non-human animal models which implies laborsome procedures and introduction of biases due to host organism-tumor interactions 9 .…”