Human Nonalcoholic Steatohepatitis on a Chip

Abstract

Nonalcoholic steatohepatitis (NASH), an advanced stage of nonalcoholic fatty liver disease (NAFLD), is a rapidly growing and global health problem compounded by the current absence of specific treatments. A major limiting factor in the development of new NASH therapies is the absence of models that capture the unique cellular structure of the liver microenvironment and recapitulate the complexities of NAFLD progression to NASH. Organ-on-a-chip platforms have emerged as a powerful approach to dynamically model diseases and test drugs. Herein, we describe a NASH-on-a-chip platform. Four main types of human primary liver cells (hepatocytes [HCs], Kupffer cells, liver sinusoidal endothelial cells, and hepatic stellate cells [HSCs]) were cocultured under microfluidic dynamics. Our chip-based model successfully recapitulated a functional liver cellular microenvironment with stable albumin and urea secretion for at least 2 weeks. Exposing liver chips to a lipotoxic environment led to gradual development of NASH phenotypic characteristics, including intracellular lipid accumulation, hepatocellular ballooning, HSC activation, and elevation of inflammatory and profibrotic markers. Further, exposure of the chip to elafibranor, a drug under study for the therapy of NASH, inhibited the development of NASH-specific hallmarks, causing an ~8-fold decrease in intracellular lipids, a 3-fold reduction in number of ballooned HCs, a significant reduction in HSC activation, and a significant decrease in the levels of inflammatory and profibrotic markers compared with controls. Conclusion: We have successfully developed a microfluidic NASH-on-a-chip platform that recapitulates the main NASH histologic endpoints in a single chip and that can emerge as a powerful noninvasive, human-relevant, in vitro platform to study disease pathogenesis and develop novel anti-NASH drugs. (Hepatology Communications 2020;0:1-17). N onalcoholic fatty liver disease (NAFLD), recently redefined as metabolic (dysfunction)-associated fatty liver disease is a widespread rising epidemic affecting more than 25% of the world's population. (1) NAFLD is now recognized as the most frequent cause of chronic liver diseases around the world and is expected to be the main cause of liver transplantation in the United States by 2030. (2) NAFLD constitutes a wide spectrum of liver pathologies starting from benign fat accumulation (hepatic steatosis) to a more complex and severe liver pathology known as nonalcoholic steatohepatitis (NASH),

Microtechnology-based in vitro models: Mimicking liver function and pathophysiology

“…5(e) ]. 113 Four types of primary cells were co-cultured with stable viability, and it was confirmed that liver-specific albumin and urea secretion function was improved. NASH was induced by exposure to FFA and LPS, and expression levels were observed by analyzing inflammatory markers MCP1, TNF-alpha, TGF-beta, and OPN.…”

“…Copyright 2021 John Wiley and Sons. 113 (f) HBV was induced in liver-on-a-chip by transfection with HBV-genome cDNA and virus genome expressed from recombinant adenovirus. Reproduced with permission from Kang et al , Biotechnol.…”

In vitro models for non-alcoholic fatty liver disease: Emerging platforms and their applications

“…Through a 21-day life-span, the system recapitulated NASH features and showed neovascularization. Freag et al. (2021) also developed an on-chip model by co-culturing primary human hepatic cell lines (hepatocytes, KC, HSC and LSEC).…”

Relevance of organ(s)-on-a-chip systems to the investigation of food-gut microbiota-host interactions

“…More recently, a model was created to use alcohol as an inductor to study biomarkers such as oxidative stress and lipid accumulation and to generate a novel in vitro quantitative readout of alcoholic liver toxicity by measuring structural changes of the bile canaliculi network (Nawroth et al 2020). There are also models for the study of NAFLD, which comprises different stages, from steatosis to steatohepatitis, cirrhosis and hepatocellular carcinoma (Gori et al 2016;Bulutoglu et al 2019;Cho et al 2021;Freag et al 2021).…”