Identification of hepatic fibrosis inhibitors through morphometry analysis of a hepatic multicellular spheroids model

• Published in: Scientific reports Vol. 11; no. 1; pp. 10931 - 12
• Main Authors: Song, Yeonhwa, Kim, Sanghwa, Heo, Jinyeong, Shum, David, Lee, Su-Yeon, Lee, Minji, Kim, A-Ram, Seo, Haeng Ran
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 25.05.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 631/154; 631/67; 692/4020; Acids; Actin; Antitumor agents; Apoptosis; Carcinoma, Hepatocellular - drug therapy; Carcinoma, Hepatocellular - metabolism; Carcinoma, Hepatocellular - pathology; Cell Line, Tumor; Colforsin - pharmacology; Drug development; Drug screening; Drug Synergism; Drugs; Endothelial cells; Epithelial-Mesenchymal Transition - drug effects; Extracellular matrix; Fibroblasts; Fibroblasts - cytology; Fibrosis; Forskolin; Hep G2 Cells; Hepatic Stellate Cells - cytology; Hepatocellular carcinoma; High-throughput screening; Human Umbilical Vein Endothelial Cells; Humanities and Social Sciences; Humans; Inflammation; Inhibitor drugs; Liver cancer; Liver Cirrhosis - drug therapy; Liver Cirrhosis - metabolism; Liver Cirrhosis - pathology; Liver Neoplasms - drug therapy; Liver Neoplasms - metabolism; Liver Neoplasms - pathology; Mesenchyme; Microenvironments; Morphometry; Multicellular tumor spheroids; multidisciplinary; Retinoic acid; Science; Science (multidisciplinary); Small Molecule Libraries - pharmacology; Sorafenib - pharmacology; Spheroids; Spheroids, Cellular; Stellate cells; Stem cells; Stromal cells; Tretinoin - pharmacology; Tumor Microenvironment
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-021-90263-x

A chronic, local inflammatory milieu can cause tissue fibrosis that results in epithelial-to-mesenchymal transition (EMT), endothelial-to-mesenchymal transition (EndMT), increased abundance of fibroblasts, and further acceleration of fibrosis. In this study, we aimed to identify potential mechanisms and inhibitors of fibrosis using 3D model-based phenotypic screening. We established liver fibrosis models using multicellular tumor spheroids (MCTSs) composed of hepatocellular carcinoma (HCC) and stromal cells such as fibroblasts (WI38), hepatic stellate cells (LX2), and endothelial cells (HUVEC) seeded at constant ratios. Through high-throughput screening of FDA-approved drugs, we identified retinoic acid and forskolin as candidates to attenuate the compactness of MCTSs as well as inhibit the expression of ECM-related proteins. Additionally, retinoic acid and forskolin induced reprogramming of fibroblast and cancer stem cells in the HCC microenvironment. Of interest, retinoic acid and forskolin had anti-fibrosis effects by decreasing expression of α-SMA and F-actin in LX2 cells and HUVEC cells. Moreover, when sorafenib was added along with retinoic acid and forskolin, apoptosis was increased, suggesting that anti-fibrosis drugs may improve tissue penetration to support the efficacy of anti-cancer drugs. Collectively, these findings support the potential utility of morphometric analyses of hepatic multicellular spheroid models in the development of new drugs with novel mechanisms for the treatment of hepatic fibrosis and HCCs.