A Drug Screening Pipeline Using 2D and 3D Patient-Derived In Vitro Models for Pre-Clinical Analysis of Therapy Response in Glioblastoma

Abstract

Glioblastoma is one of the most common and lethal types of primary brain tumor. Despite aggressive treatment with chemotherapy and radiotherapy, tumor recurrence within 6–9 months is common. To overcome this, more effective therapies targeting cancer cell stemness, invasion, metabolism, cell death resistance and the interactions of tumor cells with their surrounding microenvironment are required. In this study, we performed a systematic review of the molecular mechanisms that drive glioblastoma progression, which led to the identification of 65 drugs/inhibitors that we screened for their efficacy to kill patient-derived glioma stem cells in two dimensional (2D) cultures and patient-derived three dimensional (3D) glioblastoma explant organoids (GBOs). From the screening, we found a group of drugs that presented different selectivity on different patient-derived in vitro models. Moreover, we found that Costunolide, a TERT inhibitor, was effective in reducing the cell viability in vitro of both primary tumor models as well as tumor models pre-treated with chemotherapy and radiotherapy. These results present a novel workflow for screening a relatively large groups of drugs, whose results could lead to the identification of more personalized and effective treatment for recurrent glioblastoma.

Macrophages/Microglia in the Glioblastoma Tumor Microenvironment

“…GBOs maintain key features of GBM and can be rapidly deployed to investigate patient-specific treatment strategies. [ 110 , 111 ] In vivo, Hamilton L et al described a live imaging assay to study glioma–microglia interactions in the zebrafish brain. They believe that this model will be an important tool for drug screening and development [ 112 ].…”

Heterogeneity of Response to Iron-Based Metallodrugs in Glioblastoma Is Associated with Differences in Chemical Structures and Driven by FAS Expression Dynamics and Transcriptomic Subtypes

“…The value of individualized therapy for GBM has been recognized by the community [ 50 , 51 , 52 ], although the benefit in terms of improved patient outcomes has been hindered, in part, by cellular resistance mechanisms emerging in response to targeted therapies [ 53 ]. The identification of biomarkers of response to temozolomide [ 54 ], the chemotherapy used in the standard of care [ 4 ], has improved our knowledge of the mechanism of action of this drug, but the lack of alternative treatments for resistant (non MGMT methylated) GBMs has limited the clinical implications of this finding [ 55 ].…”

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

“…Following recent achievements in the discovery of GBOs [ 22 , 23 ], we studied this new model to evaluate its effectiveness on invasiveness of drugs and small-molecule inhibitors that have been effective on motility in 2D cultures [ 48 ], or invasiveness in simpler 3D spheroid models, yet may not appear to be effective for patients in all cases or in clinical trials. The rapid evaluation of compounds on the invasiveness of individual patient’s cells in the more heterogeneous and tissue-like context of the GBO model may reveal that such compounds are effective for some patient’s cells and not others, thus further helping to guide a personalized therapeutic approach.…”

“…This model has been shown to preserve tumor heterogeneity and complexity, to recapitulate inter-and intra-tumoral heterogeneity, and to maintain many key features of GBM, such as its cytoarchitecture and cell-cell interactions [20][21][22]. Our goal was to validate the use of GBOs in 3D pre-clinical models for precision medicine, specifically assaying invasiveness using our automated Matrigel-based invasion assay [23]. We used patient-derived tissue obtained from surgery and our precision medicine biobank [24].…”