Modelling human liver fibrosis in the context of non-alcoholic steatohepatitis using a microphysiological system

Abstract

Non-alcoholic steatohepatitis (NASH) is a common form of chronic liver disease characterised by lipid accumulation, infiltration of immune cells, hepatocellular ballooning, collagen deposition and liver fibrosis. There is a high unmet need to develop treatments for NASH. We have investigated how liver fibrosis and features of advanced clinical disease can be modelled using an in vitro microphysiological system (MPS). The NASH MPS model comprises a co-culture of primary human liver cells, which were cultured in a variety of conditions including+/− excess sugar, fat, exogenous TGFβ or LPS. The transcriptomic, inflammatory and fibrotic phenotype of the model was characterised and compared using a system biology approach to identify conditions that mimic more advanced clinical disease. The transcriptomic profile of the model was shown to closely correlate with the profile of patient samples and the model displayed a quantifiable fibrotic phenotype. The effects of Obeticholic acid and Elafibranor, were evaluated in the model, as wells as the effects of dietary intervention, with all able to significantly reduce inflammatory and fibrosis markers. Overall, we demonstrate how the MPS NASH model can be used to model different aspects of clinical NASH but importantly demonstrate its ability to model advanced disease with a quantifiable fibrosis phenotype.

Src kinase as a potential therapeutic target in non‐alcoholic and alcoholic steatohepatitis

“…We treated 7‐day spheroids with LPS, and incubated with KX2‐391 for 7 days, and found that treatment with LPS increased collagen I expression and macrophage activation marker, MHC‐II (major histocompatibility complex class II) expression as well as steatosis assessed by oil‐red‐o staining, and all these markers were reduced following Src inhibition (Figure S8B). However, it is important to stress that LPS only induces pathways associated with inflammatory signalling therefore NASH‐relevant cues such as fructose, free fatty acids, cholesterol and TGFβ in combination with LPS could be supplemented to model different aspects of clinical NASH 45 …”

“…However, it is important to stress that LPS only induces pathways associated with inflammatory signalling therefore NASHrelevant cues such as fructose, free fatty acids, cholesterol and TGFβ in combination with LPS could be supplemented to model different aspects of clinical NASH. 45…”

3D multicellular systems in disease modelling: From organoids to organ-on-chip

“…Thus, the use of 3D multicellular systems is highly beneficial for modelling this group of diseases. For instance, 3D multicellular liver models have been previously shown to accurately model the cellular phenotypes associated with non-alcoholic steatohepatitis (NASH), a metabolic disease affecting the liver ( McCarron et al, 2019 ; Ouchi et al, 2019 ; Duan et al, 2020 ; Lee-Montiel et al, 2020 ; Ramli et al, 2020 ; Yang et al, 2020 ; Daou et al, 2021 ; Freag et al, 2021 ; Kostrzewski et al, 2021 ; Slaughter et al, 2021 ). Daou et al (2021) established a liver 3D multicellular system comprising a primary human hepatocyte (PHH) model of lipotoxicity, a primary human macrophage (PHM) model, and a primary human hepatic stellate cell (HSC) model.…”

Microfluidic Organ-on-a-Chip Devices for Liver Disease Modeling In Vitro

“…Similarly, Suurmond et al induced steatosis in spheroid cells generated using AggreWell plates and explored the feedback signaling between them in the pathophysiology of NASH [ 62 ]. The three-culture spheroid microphysiological platform was used to induce the NAFLD with progression to NASH by measuring the deposition of the collagen and markers of fibrotic tissue [ 63 ]. The increase of inflammatory cytokines and activation of stellate cells were correlated with transcriptomic analysis and gene expression.…”

“…The increase of inflammatory cytokines and activation of stellate cells were correlated with transcriptomic analysis and gene expression. Moreover, the authors investigated the reversibility of the condition by evaluating the positive effect of treatment drugs in an ongoing study [ 63 ].…”