Intracellular Proteomics and Extracellular Vesiculomics as a Metric of Disease Recapitulation in 3D Bioprinted Aortic Valve Arrays

ABSTRACT In calcific aortic valve disease (CAVD), mechanosensitive valvular cells respond to fibrosis- and calcification-induced tissue stiffening, further driving pathophysiology. No pharmacotherapeutics are available to treat CAVD, due to the lack of: 1) appropriate experimental models that recapitulate this complex environment; and 2) benchmarking novel engineered AV-model performance. We established a biomaterial-based CAVD model mimicking the biomechanics of the human AV disease-prone fibrosa layer, 3D-bioprinted into 96-well arrays. LC-MS/MS analyses probed the cellular proteome and vesiculome to compare the 3D-bioprinted model vs. traditional 2D monoculture, against human CAVD tissue. The 3D-bioprinted model highly recapitulated the CAVD cellular proteome (94% vs. 70% of 2D proteins). Integration of cellular/vesicular datasets identified known and novel proteins ubiquitous to AV calcification. This study explores how 2D vs. 3D-bioengineered systems recapitulate unique aspects of human disease, positions multi-omics as a novel technique for the evaluation of high throughput-based bioengineered model systems and potentiates future drug discovery.