title: High-throughput investigation of endothelial-to-mesenchymal transformation (EndMT) with combinatorial cellular microarrays.
abstract: In the developing heart, a specific subset of endocardium undergoes an endothelial-to-mesenchymal transformation (EndMT) thus forming nascent valve leaflets. Extracellular matrix (ECM) proteins and growth factors (GFs) play important roles in regulating EndMT but the combinatorial effect of GFs with ECM proteins is less well understood. Here we use microscale engineering techniques to create single, binary, and tertiary component microenvironments to investigate the combinatorial effects of ECM proteins and GFs on the attachment and transformation of adult ovine mitral valve endothelial cells to a mesenchymal phenotype. With the combinatorial microenvironment microarrays, we utilized 60 different combinations of ECM proteins (Fibronectin, Collagen I, II, IV, Laminin) and GFs (TGF-β1, bFGF, VEGF) and were able to identify new microenvironmental conditions capable of modulating EndMT in MVECs. Experimental results indicated that TGF-β1 significantly upregulated the EndMT while either bFGF or VEGF downregulated EndMT process markedly. Also, ECM proteins could influence both the attachment of MVECs and the response of MVECs to GFs. In terms of attachment, fibronectin is significantly better for the adhesion of MVECs among the five tested proteins. Overall collagen IV and fibronectin appeared to play important roles in promoting EndMT process. Great consistency between macroscale and microarrayed experiments and present studies demonstrates that high-throughput cellular microarrays are a promising approach to study the regulation of EndMT in valvular endothelium. Biotechnol. Bioeng. 2016;113: 1403-1412. © 2015 Wiley Periodicals, Inc.
authors.affiliation: School of Engineering, University of British Columbia, Kelowna, BC, V1V1V7, Canada.
authors.forename: Zongjie
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Institute of Molecular Health Sciences, ETH Zürich, Zürich, Switzerland.|Center for Biomedical Engineering, Brigham and Women's Hospital, Harvard Medical School, Cambridge, Massachusetts.|Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, Massachusetts.
authors.forename: Blaise
authors.initials: B
authors.lastname: Calpe
authors.affiliation: Center for Biomedical Engineering, Brigham and Women's Hospital, Harvard Medical School, Cambridge, Massachusetts.|Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, Massachusetts.|Institute of Bioengineering, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
authors.forename: Jalil
authors.initials: J
authors.lastname: Zerdani
authors.affiliation: Department of Mathematics and Statistics, University of British Columbia, Kelowna, BC, Canada.
authors.forename: Youngsang
authors.initials: Y
authors.lastname: Lee
authors.affiliation: Center for Biomedical Engineering, Brigham and Women's Hospital, Harvard Medical School, Cambridge, Massachusetts.|Division of Mechanical Design Engineering, Chonbuk National University, Jeonjoo, Republic of Korea.|Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, 02139.
authors.forename: Jonghyun
authors.initials: J
authors.lastname: Oh
authors.affiliation: Center for Biomedical Engineering, Brigham and Women's Hospital, Harvard Medical School, Cambridge, Massachusetts.|Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, 02139.|Department of Bioindustrial Technologies, College of Animal Bioscience and Technology, Konkuk University, Hwayang-dong, Kwangjin-gu, Seoul, Republic of Korea.
authors.forename: Hojae
authors.initials: H
authors.lastname: Bae
authors.affiliation: Center for Biomedical Engineering, Brigham and Women's Hospital, Harvard Medical School, Cambridge, Massachusetts. alik@rics.bwh.harvard.edu.|Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, Massachusetts. alik@rics.bwh.harvard.edu.|Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, 02139. alik@rics.bwh.harvard.edu.|Department of Physics, King Abdulaziz University, Jeddah 21569, Saudi Arabia. alik@rics.bwh.harvard.edu.
authors.forename: Ali
authors.initials: A
authors.lastname: Khademhosseini
authors.affiliation: School of Engineering, University of British Columbia, Kelowna, BC, V1V1V7, Canada. keekyoung.kim@ubc.ca.|Center for Biomedical Engineering, Brigham and Women's Hospital, Harvard Medical School, Cambridge, Massachusetts. keekyoung.kim@ubc.ca.|Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, 02139. keekyoung.kim@ubc.ca.
authors.forename: Keekyoung
authors.initials: K
authors.lastname: Kim
mesh_terms: D000818:Animals; D054338:Cell Transdifferentiation / Q000502:physiology*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D006351:Heart Valves / Q000166:cytology; D057166:High-Throughput Screening Assays / Q000379:methods*; D036341:Intercellular Signaling Peptides and Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D051381:Rats; D013552:Swine; D046888:Tissue Array Analysis / Q000379:methods*
chemical_list: D016326:Extracellular Matrix Proteins; D036341:Intercellular Signaling Peptides and Proteins
keywords: endothelial-to-mesenchymal transformation; heart valve; high-throughput; microarray