High-throughput investigation of endothelial-to-mesenchymal transformation (EndMT) with combinatorial cellular microarrays

• Published in: Biotechnology and bioengineering 2016-07, Vol.113 (7), p.1403-1412
• Main Authors: Wang, Zongjie, Calpe, Blaise, Zerdani, Jalil, Lee, Youngsang, Oh, Jonghyun, Bae, Hojae, Khademhosseini, Ali, Kim, Keekyoung
• Format: Article
• Language: English
• Subjects: Adults; Animals; Attachment; Bioengineering; Cell Transdifferentiation - physiology; Cellular; Cellular biology; Combinatorial analysis; Electrochemical machining; Endothelial Cells - cytology; Endothelial Cells - metabolism; endothelial-to-mesenchymal transformation; Extracellular Matrix Proteins - analysis; Extracellular Matrix Proteins - genetics; Extracellular Matrix Proteins - metabolism; Fibronectin; Heart; heart valve; Heart Valves - cytology; high-throughput; High-Throughput Screening Assays - methods; Intercellular Signaling Peptides and Proteins - analysis; Intercellular Signaling Peptides and Proteins - genetics; Intercellular Signaling Peptides and Proteins - metabolism; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - metabolism; microarray; Proteins; Rats; Swine; Tissue Array Analysis - methods; Transformations
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.25905

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. In the developing heart, a subset of endocardium undergoes an endothelial‐to‐mesenchymal transformation (EndMT). The authors use microscale techniques to create single, binary, and tertiary component microenvironments to investigate the combinatorial effects of Extracellular matrix (ECM) proteins and growth factors (GFs) on the attachment and transformation of adult ovine mitral valve endothelial cells (MVECs) to a mesenchymal phenotype. The authors utilized 60 different combinations of ECM proteins and GFs and were able to identify new microenvironmental conditions capable of modulating EndMT in MVECs.