Design and Biological Activity of a Novel Urea Compound Against Glioblastoma [scRNAseq_MXC-017_374_in_vivo]
Despite advances in surgery and radiotherapy, glioblastoma (GBM) remains the deadliest adult brain cancer with unacceptable low median survival rates. Targeted therapies and biologics have failed to improve outcome and the standard-of-care against GBM has not changed in two decades. GBM, like many solid tumors are thought to be organized hierarchically with a small number of radiation- and chemotherapy-resistant glioma stem cells (GSCs) producing more differentiated progeny and can repopulate tumors after sublethal treatment.In this study we sought to develop novel compounds that cross the blood-brain barrier (BBB), target existing GSCs and interfere with glioma cell plasticity to prevent the induction of GSCs in response to irradiation. We show that novel urea compounds target existing GSCs, prevent the radiation-induced conversion of non-stem glioma cells into GSCs and prolong the median survival in mouse models of GBM with minimal normal tissue toxicity. Overall design: 300,000 HK-374 cells, engineered to express GFP and firefly luciferase, were implanted intracranially and allowed to engraft for one week. Engraftment was verified by bioluminescent imaging, after which mice received either a single 4 Gy dose of radiation alone or radiation followed by MXC-017 treatment (5 i.p. injections per week, 50 mg/kg) for two weeks. Tumors were then harvested and dissociated into single cells using the Miltenyi Brain Tumor Dissociation Kit (#130-095-942), followed by mouse cell depletion (Mouse Cell Depletion Kit, #130-104-694, Miltenyi). Cells from 2-3 mice were pooled and subjected for sequencing.