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bioRxiv · 10.1101/2023.06.22.546103

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

Clift, C. L.·Blaser, M. C.·Gerrits, W.·Turner, M. E.·Sonawane, A. C.·Pham, T.·Andresen, J. L.·Fenton, O. S.·Grolman, J. M.·Buffolo, F.·Schoen, F. J.·Hjortnaes, J.·Muehlschlegel, J. D.·Mooney, D. D.·Aikawa, M.·Singh, S. A.·Langer, R.·Aikawa, E.
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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.

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Clift, C. L., Blaser, M. C., Gerrits, W., Turner, M. E., Sonawane, A. C., Pham, T., Andresen, J. L., Fenton, O. S., Grolman, J. M., Buffolo, F., Schoen, F. J., Hjortnaes, J., Muehlschlegel, J. D., Mooney, D. D., Aikawa, M., Singh, S. A., Langer, R., Aikawa, E.. 2023-06-25. Intracellular Proteomics and Extracellular Vesiculomics as a Metric of Disease Recapitulation in 3D Bioprinted Aortic Valve Arrays. https://doi.org/10.1101/2023.06.22.546103

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