Three-Dimensional in vitro Models of Healthy and Tumor Brain Microvasculature for Drug and Toxicity Screening.

Marie Piantino,Agathe Figarol,Michiya Matsusaki

doi:10.3389/ftox.2021.656254

Abstract

Tissue vascularization is essential for its oxygenation and the homogenous diffusion of nutrients. Cutting-edge studies are focusing on the vascularization of three-dimensional (3D) in vitro models of human tissues. The reproduction of the brain vasculature is particularly challenging as numerous cell types are involved. Moreover, the blood-brain barrier, which acts as a selective filter between the vascular system and the brain, is a complex structure to replicate. Nevertheless, tremendous advances have been made in recent years, and several works have proposed promising 3D in vitro models of the brain microvasculature. They incorporate cell co-cultures organized in 3D scaffolds, often consisting of components of the native extracellular matrix (ECM), to obtain a micro-environment similar to the in vivo physiological state. These models are particularly useful for studying adverse effects on the healthy brain vasculature. They provide insights into the molecular and cellular events involved in the pathological evolutions of this vasculature, such as those supporting the appearance of brain cancers. Glioblastoma multiform (GBM) is the most common form of brain cancer and one of the most vascularized solid tumors. It is characterized by a high aggressiveness and therapy resistance. Current conventional therapies are unable to prevent the high risk of recurrence of the disease. Most of the new drug candidates fail to pass clinical trials, despite the promising results shown in vitro. The conventional in vitro models are unable to efficiently reproduce the specific features of GBM tumors. Recent studies have indeed suggested a high heterogeneity of the tumor brain vasculature, with the coexistence of intact and leaky regions resulting from the constant remodeling of the ECM by glioma cells. In this review paper, after summarizing the advances in 3D in vitro brain vasculature models, we focus on the latest achievements in vascularized GBM modeling, and the potential applications for both healthy and pathological models as platforms for drug screening and toxicological assays. Particular attention will be paid to discuss the relevance of these models in terms of cell-cell, cell-ECM interactions, vascularization and permeability properties, which are crucial parameters for improving in vitro testing accuracy.

Highlights

Importance of Vascularization in 3D EngineeringFor decades, the construction of vascularized tissue has been a major challenge in 3D tissue engineering for non-animal alternatives
Glioblastoma multiform (GBM) is one of the most common forms of brain cancer in adults, and one of the deadliest brain tumors, with a median survival of 12 months with appropriate treatments. It is one of the most vascularized brain cancers, and is associated with a high remodeling of the extracellular matrix (ECM) There has been intensive research dedicated to modeling the characteristic features of GBM in order to understand its impact on brain vascularization, the regulation of the angiogenic signaling pathways, as microvascular proliferation is a hallmark of GBM (Hardee and Zagzag, 2012; Rodriguez et al, 2012)
Yi et al developed a bioprinted GBM on-a-chip supported by a brain-derived ECM using patient-derived GBM cancer cells in co-culture with Human Umbilical Vein Endothelial Cells (HUVECs), with each cell type compartmentalized in specific regions of the model (Yi et al, 2019)

Summary

Introduction

Importance of Vascularization in 3D EngineeringFor decades, the construction of vascularized tissue has been a major challenge in 3D tissue engineering for non-animal alternatives. Cho et al proposed a fully human model composed of either immortalized cell line ECs (hCMEC/D3) or primary microvascular ECs, in co-culture with primary ACs and PCs. Brain ECs demonstrated barrier function properties, including tight junctions formation and efflux transporter activity, as assessed by the high expression level of P-gp efflux pump on the surface of the spheroid.

Results

Conclusion