Development and validation of an in vitro 3D model of NASH with severe fibrotic phenotype

• Published in: American journal of translational research Vol. 11; no. 3; pp. 1531 - 1540
• Main Authors: Mukherjee, Sumanta, Zhelnin, Leonid, Sanfiz, Anthony, Pan, Jie, Li, Zhuyin, Yarde, Melissa, McCarty, Jean, Jarai, Gabor
• Format: Journal Article
• Language: English
• Published: United States e-Century Publishing Corporation 01.01.2019
• Subjects: Original; liver micro tissue; fibrosis; NASH; ASK-1
• ISSN: 1943-8141; 1943-8141

Nonalcoholic steatohepatitis represents a significant and rapidly growing unmet medical need. The development of novel therapies has been hindered in part, by the limitations of existing preclinical models. There is a strong need for physiologically relevant and liver fibrosis models that are characterized by better translational predictability. In this study, we used the InSphero 3D InSight three-dimensional (3D) human liver microtissue (3D-hLMT) system prepared by co-culturing primary human hepatocytes with hepatic stellate cells, Kupffer cells and endothelial cells to develop a model of NASH with a severe fibrotic phenotype. In our model, palmitic acid (PA) induced a robust proinflammatory and profibrogenic phenotype in the 3D-hLMT. PA significantly increased several markers of the inflammatory and profibrotic process including gene expression of collagens, , tissue inhibitor of matrix metalloprotease 1 ( ) and the stellate cell activation marker as well as secreted CXCL8 (IL8) levels. We also observed TGFβ pathway activation, increase in active collagen synthesis and significant overall increase in tissue damage in the 3D-hLMTs. Immunohistochemistry analysis demonstrated the upregulation of collagen, cleaved caspase 3 as well as of the PDGFRβ protein. We further validated the model using a phase 3 clinical compound, GS-4997, an apoptosis signal-regulating kinase 1 (ASK-1) inhibitor and showed that GS-4997 significantly decreased PA induced profibrotic and proinflammatory response in the 3D-hLMTs with decreases in apoptosis and stellate cell activation in the microtissues. Taken together we have established and validated an 3D-hLMT NASH model with severe fibrotic phenotype that can be a powerful tool to investigate experimental compounds for the treatment of NASH.