Table 2.

Uses of in vitro NVU models to study CNS disease pathogenesis

Disease	In vitro NVU Model	Major Findings	Reference
Alzheimer’s Disease	Microfluidic NVU model with hCMEC/D3 and control or AD ReN human neural progenitor cell cells	BBB dysfunctions were present in AD models, including increased permeability, reduced tight junction protein expression, increased MMP2 and reactive oxygen species and deposition of amyloid at the vascular endothelium.	[46]
	Isogenic APOE3 and APOE4 iPSC-derived vascular organoid model	Dysregulation of calcineurin–NFAT signaling and APOE in pericyte-like mural cells induced APOE4-associated pathology.	[19]
	Three-dimensional bioengineered vessel NVU model	ApoE and HDL synergized to facilitate Aβ transport across the endothelial cell barrier. ApoE4 was less efficient than ApoE2 in promoting Aβ transport.	[56]
	iPSC-derived Transwell NVU model	Aβ and neuroinflammation signals increased IgG uptake and transport in the model.	[57]
	Mouse brain endothelial cell Transwell model	Aβ1–42 induced tight junction damage and BBB leakage in bEnd.3 Transwell model. RAGE played an important role in the process.	[58]
	Primary mouse brain capillary endothelial cell Transwell model	Basolateral recombinant ApoJ and apical ApoA1 facilitated the transport of basolateral Aβ1–40 in this model.	[59]
	Rat brain microvascular endothelial cells	Exposure to Aβ25–35 disrupted tight junctions, increased BBB permeability, decreased cell viability in this model.	[60]
	hCMEC/D3 Transwell model	hCMEC/D3 cells had limited utility in studying Aβ trafficking due to the low barrier tightness of this model.	[61]
Parkinson’s Disease	Microfluidic NVU model with iPSCdopaminergic neurons, iPSC-derived BMEC-like cells, and primary human astrocytes, microglia, and pericytes	α-Synuclein fibril treatment induced key aspects of Parkinson’s disease phenotypes, including accumulation of pSer129-αSyn, mitochondrial impairment, neuroinflammation, and increased BBB permeability.	[48]
	hCMEC/D3 Transwell model	α-Synuclein pre-formed fibrils impaired tight junction protein expression in endothelial cells.	[47]
	Rat cerebral microvessel endothelial cells and C6 astroglial cells Transwell model	PD drug FLZ was effluxed by P-gp in rat cerebral microvessel endothelial cells.	[62]
	Primary rat brain endothelial cells and pericytes Transwell model	Pericytes were more sensitive to monomeric α-synuclein than endothelial cells regarding release of inflammatory cytokines and MMP-9 in this model.	[63]
Viral Infection	Microfluidic NVU model with hCMEC/D3 cells	SARS-CoV-2 spike protein S1 triggered a pro-inflammatory response and promotes loss of barrier integrity in this model.	[49]
	iPSC-pericyte-likecell-containing cortical organoid	Pericyte-like cells integrated into the cortical organoid were infected with SARS-CoV-2, and served as viral replication hubs.	[64]
	hCMEC/D3 Transwell model	Zika virus infected hCMEC/D3 cells without disrupting BBB permeability and tight junction protein expression, and the virus was subsequently released on the brain side.	[65]
	iPSC-derived Transwell NVU model	Zika virus infected iPSC-derived BMEC-like cells without disrupting BBB permeability and tight junction protein expression, and was subsequently released on the brain side.	[50]
	Primary human brain microvascular endothelial cells Transwell model	West Nile virus infected primary human BMECs, leading to increased permeability, increased leukocyte adhesion and transmigration across the in vitro model.	[66]