Phenotypic screening in Organ-on-a-Chip systems: a 1537 kinase inhibitor library screen on a 3D angiogenesis assay

Abstract

Modern drug development increasingly requires comprehensive models that can be utilized in the earliest stages of compound and target discovery. Here we report a phenotypic screening exercise in a high-throughput Organ-on-a-Chip setup. We assessed the inhibitory effect of 1537 protein kinase inhibitors in an angiogenesis assay. Over 4000 micro-vessels were grown under perfusion flow in microfluidic chips, exposed to a cocktail of pro-angiogenic factors and subsequently exposed to the respective kinase inhibitors. Efficacy of compounds was evaluated by reduced angiogenic sprouting, whereas reduced integrity of the main micro-vessel was taken as a measure for toxicity. The screen yielded 53 hits with high anti-angiogenicity and low toxicity, of which 44 were previously unassociated with angiogenic pathways. This study demonstrates that Organ-on-a-Chip models can be screened in high numbers to identify novel compounds and targets. This will ultimately reduce bias in early-stage drug development and increases probability to identify first in class compounds and targets for today’s intractable diseases.

Development of a cancer metastasis-on-chip assay for high throughput drug screening

“…Previous studies have used this platform to conduct high throughput analysis for T cell extravasation under flow ( 48 ), endothelial cell permeability ( 31 ), and the integrity of intestinal tract epithelium ( 49 ). The same platform was employed to screen small compound libraries on retinal endothelial permeability ( 32 ) and sprouting angiogenesis ( 50 ). Our current model incorporates many relevant features of tumor microenvironment, including perfused microvessel, ECM, and stromal fibroblasts.…”

Reconstitution of human tissue barrier function for precision and personalized medicine

“…127 Additionally, hydrogel-based tissue barrier chips have been developed with models incorporating 2 wells each for inlet and outlet of the hydrogel channel and the medium supplying channel, resulting in a 4-well model in 384 well plate format. 94,124 There are also versions with two side medium channels, constituting a 6-well model 51,128 or 9-well model. 31,103…”

“…Furthermore, in 2023, the semi-automated high-throughput screening of the inhibitory effects of 1537 protein kinase inhibitors was demonstrated in an angiogenesis model. 128 Using an automated liquid handler, they constructed over 4000 endothelial barrier models, exposing them to the respective kinase inhibitors. While reagent preparations and plate transfers were carried out manually, 96.5% (3987) of the 4130 chips passed the quality control of visual inspection.…”