1. Cirrhotic Human Liver Extracellular Matrix 3D Scaffolds Promote Smad-Dependent TGF-β1 Epithelial Mesenchymal Transition
- Author
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Benjamin Robinson, Katrin Böttcher, Massimo Pinzani, Massimo Malagó, Giusi Marrone, Paolo De Coppi, Claire Crowley, Ana Levi, Giuseppe Mazza, Armando del Río Hernández, Diana Julie Leeming, Kessarin Thanapirom, Gabriele Spoletini, Jasminka Godovac-Zimmermann, Morten A. Karsdal, Mark Crawford, Lisa Longato, Walid Al-Akkad, Maria Giovanna Vilia, Andrew R. Hall, Tu Vinh Luong, L. Frenguelli, Domenico Tamburrino, Andrea Telese, Krista Rombouts, Benedetta Lombardi, and Martina Marrali
- Subjects
0301 basic medicine ,Liver Cirrhosis ,Proteomics ,Cell ,MICROENVIRONMENT ,Smad Proteins ,SMAD ,decellularized extracellular matrix (dECM) ,Smad2 Protein ,Transforming growth factor beta1 (TGF-beta 1) ,Extracellular matrix ,hepatocellular carcinoma (HCC) ,0302 clinical medicine ,Tissue engineering ,HEPATOCELLULAR-CARCINOMA ,Phosphorylation ,lcsh:QH301-705.5 ,Decellularization ,biology ,Tissue Scaffolds ,Chemistry ,TGF-BETA ,General Medicine ,Immunohistochemistry ,hepatocellular carcinoma cells ,3. Good health ,Cell biology ,Extracellular Matrix ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,tissue engineering ,3D ECM scaffolds ,Collagen ,Life Sciences & Biomedicine ,Transforming growth factor beta1 (TGF-β1) ,Signal Transduction ,Carcinoma, Hepatocellular ,Epithelial-Mesenchymal Transition ,STROMA ,FIBRONECTIN ,Bioengineering ,HEPATIC STELLATE CELLS ,decellularized human liver ,Article ,Transforming Growth Factor beta1 ,03 medical and health sciences ,proteomics ,medicine ,Galunisertib ,Humans ,Epithelial–mesenchymal transition ,tumor microenvironment (TME) ,Smad3 Protein ,Science & Technology ,Cell Biology ,Fibronectin ,030104 developmental biology ,lcsh:Biology (General) ,TISSUE ,3-dimensional (3D) platform ,biology.protein - Abstract
An altered liver microenvironment characterized by a dysregulated extracellular matrix (ECM) supports the development and progression of hepatocellular carcinoma (HCC). The development of experimental platforms able to reproduce these physio-pathological conditions is essential in order to identify and validate new therapeutic targets for HCC. The aim of this work was to validate a new in vitro model based on engineering three-dimensional (3D) healthy and cirrhotic human liver scaffolds with HCC cells recreating the micro-environmental features favoring HCC. Healthy and cirrhotic human livers ECM scaffolds were developed using a high shear stress oscillation-decellularization procedure. The scaffolds bio-physical/bio-chemical properties were analyzed by qualitative and quantitative approaches. Cirrhotic 3D scaffolds were characterized by biomechanical properties and microarchitecture typical of the native cirrhotic tissue. Proteomic analysis was employed on decellularized 3D scaffolds and showed specific enriched proteins in cirrhotic ECM in comparison to healthy ECM proteins. Cell repopulation of cirrhotic scaffolds highlighted a unique up-regulation in genes related to epithelial to mesenchymal transition (EMT) and TGF&beta, signaling. This was also supported by the presence and release of higher concentration of endogenous TGF&beta, 1 in cirrhotic scaffolds in comparison to healthy scaffolds. Fibronectin secretion was significantly upregulated in cells grown in cirrhotic scaffolds in comparison to cells engrafted in healthy scaffolds. TGF&beta, 1 induced the phosphorylation of canonical proteins Smad2/3, which was ECM scaffold-dependent. Important, TGF&beta, 1-induced phosphorylation of Smad2/3 was significantly reduced and ECM scaffold-independent when pre/simultaneously treated with the TGF&beta, R1 kinase inhibitor Galunisertib. In conclusion, the inherent features of cirrhotic human liver ECM micro-environment were dissected and characterized for the first time as key pro-carcinogenic components in HCC development.
- Published
- 2019