Predicting In Vivo Payloads Delivery using a Blood-brain Tumor-barrier in a Dish

Abstract

Highly selective by nature, the blood-brain barrier (BBB) is essential for the brain homeostasis in physiological conditions. However, in the context of brain tumors, the molecular selectivity of BBB also shields the neoplastic cells by blocking the delivery of peripherally administered chemotherapies. The development of novel drugs (including nanoparticles) targeting malignant brain tumors ideally requires the use of preclinical animal models to study the drug's transcytosis and antitumor efficacy. In order to comply with the 3R principle (refine, reduce, and replace) to reduce the number of laboratory animals in experimental setup and perform the high-throughput screening of a large library of antitumor agents, we developed a reproducible in vitro human and murine mimic of the blood-brain tumor-barrier (BBTB) using three-layered cultures of endothelial cells, astrocytes, and patient-derived glioblastoma spheres. For higher scalability and reproducibility, commercial cell lines or immortalized cells have been used in tailored conditions to allow the formation of a barrier resembling the actual BBB. Here we describe a protocol to obtain a BBTB mimic by culturing endothelial cells in contact with astrocytes at specific cell densities on inserts. This BBTB mimic can be used, for instance, for the quantification and confocal imaging of the nanoparticle passage through the endothelial and astrocytic barriers, in addition to the evaluation of the tumor cell targeting within the same assay. Moreover, we show that the obtained data can be used to predict the behavior of nanoparticles in preclinical animal models. In a broader perspective, this in vitro model could be adapted to other neurodegenerative diseases for the determination of the passage of new therapeutic molecules through the BBB and/or be supplemented with brain organoids to directly evaluate the efficacy of drugs.

Monotherapy efficacy of blood–brain barrier permeable small molecule reactivators of protein phosphatase 2A in glioblastoma

“…We, therefore, started by investigating the in vitro BBB passage of NZ-8-061 (a.k.a DT-061) that has been widely used in cancers outside the CNS ( Sangodkar et al , 2017 ; Kauko et al , 2018 ; McClinch et al , 2018 ). Quantified by HPLC-MS/MS, NZ-8-061 was found to cross the artificial BBB ( Le Joncour et al , 2019 b ), consisting of murine brain microcapillary endothelial cells and astrocytes ( Fig. 1B and C ).…”

Sunitinib-Containing Carborane Pharmacophore with the Ability to Inhibit Tyrosine Kinases Receptors FLT3, KIT and PDGFR-β, Exhibits Powerful In Vivo Anti-Glioblastoma Activity

“…Secondly, as displayed at Table 4 , compound 1 has proven not to be mutagenic, using an Ames test with two different S. typhimurium strains, another desirable characteristic that would act in favor of this compound as a drug [ 47 ]. Finally, and thinking in the potential use of 1 as anti-glioblastoma agent, we studied its ability to cross the brain blood barrier (BBB) using a reported artificial endothelial-cell model [ 48 , 49 , 50 , 51 ]. According to the results, compound 1 has the ability to cross the BBB, as ≈3-times more compound translocates across the artificial BBB after hybrid 1 exposure in the feeding side of the membrane for 24 h at room temperature in the dark and without stirring ( Table 4 ).…”

Circumventing drug treatment? Polyethylenimine functionalized nanoparticles inherently and selectively kill patient-derived glioblastoma stem cells

“…Additionally, to deduce the potential of this mechanism to be exploited in a therapeutic setting, we showed successful penetration of PEI-MSNs through BBB models both in vitro and in vivo. Furthermore, the in vitro blood-brain tumor-barrier (BBTB) model [37] confirmed accumulation of PEI-MSNs in the lysosomes of BT-12 GSC. Potentially, this discovery of the inherent role of PEI-MSNs in selectively eradicating otherwise highly resistant GSCs presents a novel vulnerability to exploit for brain cancer (GB) treatment.…”