Intracellular proteomics and extracellular vesiculomics as a metric of disease recapitulation in 3D-bioprinted aortic valve arrays

Author:

Clift Cassandra L.1ORCID,Blaser Mark C.1ORCID,Gerrits Willem12,Turner Mandy E.1ORCID,Sonawane Abhijeet1ORCID,Pham Tan1,Andresen Jason L.34,Fenton Owen S.45ORCID,Grolman Joshua M.678ORCID,Campedelli Alesandra1ORCID,Buffolo Fabrizio19ORCID,Schoen Frederick J.10ORCID,Hjortnaes Jesper11ORCID,Muehlschlegel Jochen D.12ORCID,Mooney David J.67ORCID,Aikawa Masanori113,Singh Sasha A.1ORCID,Langer Robert3141516ORCID,Aikawa Elena113ORCID

Affiliation:

1. Division of Cardiovascular Medicine, Department of Medicine, Center for Interdisciplinary Cardiovascular Sciences, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115, USA.

2. Department of Cardiothoracic Surgery, University Medical Center Utrecht, Utrecht, Netherlands.

3. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.

4. Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.

5. Division of Pharmacoengineering and Molecular Pharmaceutics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

6. John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02134, USA.

7. Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA 02115, USA.

8. Materials Science and Engineering, The Technion-Israel Institute of Technology, Haifa, Israel.

9. Division of Internal Medicine and Hypertension Unite, Department of Medical Sciences, University of Torin, Turin, Italy.

10. Department of Pathology, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA 02115, USA.

11. Department of Cardiothoracic Surgery, Leiden University Medical Center (LUMC), Leiden, Netherlands.

12. Department of Anesthesiology, Perioperative and Pain Medicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115, USA.

13. Division of Cardiovascular Medicine, Department of Medicine, Center for Excellence in Vascular Biology, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115, USA.

14. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02142, USA.

15. Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, MA 02142, USA.

16. Harvard and MIT Division of Health Science and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.

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 because of the paucity of (i) appropriate experimental models that recapitulate this complex environment and (ii) benchmarking novel engineered aortic valve (AV)–model performance. We established a biomaterial-based CAVD model mimicking the biomechanics of the human AV disease-prone fibrosa layer, three-dimensional (3D)–bioprinted into 96-well arrays. Liquid chromatography–tandem mass spectrometry analyses probed the cellular proteome and vesiculome to compare the 3D-bioprinted model versus traditional 2D monoculture, against human CAVD tissue. The 3D-bioprinted model highly recapitulated the CAVD cellular proteome (94% versus 70% of 2D proteins). Integration of cellular and vesicular datasets identified known and unknown proteins ubiquitous to AV calcification. This study explores how 2D versus 3D-bioengineered systems recapitulate unique aspects of human disease, positions multiomics as a technique for the evaluation of high throughput–based bioengineered model systems, and potentiates future drug discovery.

Publisher

American Association for the Advancement of Science (AAAS)

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