Abstract

A 3D Bioprinted Neurovascular Unit Model for Glioblastoma Brain Tumor Formation and Drug Screening

Yen-Ting Tung, Research Fellow, National Center for Advancing Translational Sciences (NCATS)

A bioprinted 3D neurovascular unit (NVU) model was developed to study glioblastoma (GBM) tumor formation in a brain-like microenvironment. This NVU model was bioprinted on a 96-well plate format to enable high-through-put screening for anti-GBM therapies. The bioprinted NVU model included human primary astrocytes, pericytes and GFP-tagged brain microvascular endothelial cells. The patient-derived glioblastoma cells, JHH520, were tagged with mCherry and added into the NVU model. We used confocal high content imaging to quantitate formation of a vascular network by vasculogenesis and angiogenesis and GBM tumor growth with GFP expressing endothelial cells and mCherry GBM cells, respectively. Extensive validation of the NVU-GBM tissue model included immunostaining for deposited brain relevant extra cellular matrix components, including collagen IV, laminin, and fibronectin; single cell RNA sequencing to establish clinically relevant transcriptomics changes; and secretion of NVU and GBM-relevant cytokines. During the formation of the NVU, we observed physiologically relevant changes in gene expression and cytokines secretion associated with fibrosis/wound healing/angiogenesis. This engineered NVU-GBM model was used to test the antitumor effects of 18 drugs to assess its pharmacological relevance. This tissue biofabrication approach demonstrated here is versatile and modular, and will enable the inclusion of patient tumor cells for personalized medicine.