in vivo MASH (NASH) model, Liver on a Chip, MASH Biomarkers
Translational Research service through Humanized Disease Models
Overview： in vivo MASH (NASH) model
Metabolic Dysfunction Associated Steatohepatitis (MASH) is one of the most prevalent chronic liver diseases, which is closely associated with obesity, insulin resistance and dyslipidemia. These important features therefore need to be reflected in a preclinical model as well. Based on 25 years of research on translational metabolic disease models, TNO has developed the Ldlr-/-.Leiden MASH mouse model that accurately mimics the etiology and pathology of MASH and fibrosis in humans. By using a high-fat diet, with a macronutrient composition comparable to human diets (e.g., without added cholesterol), the Ldlr-/-.Leiden MASH mouse model develops obesity, insulin resistance, adipose tissue inflammation, increased gut permeability with altered microbiota composition, and MASH with bridging fibrosis [F3 stage].
Features：the Ldlr-/-.Leiden MASH mouse model
● Translational disease induction and clinical phenotype
● Severe obesity (with adipose tissue inflammation)
● Insulin resistance
● Hyperlipidemia (humanized lipoprotein profiles), high triglycerides, high (V)LDL, low HDL)
● Translational histology: steatosis, inflammatory aggregates and bridging fibrosis (up to F3)
● Atherosclerosis development
● Translational underlying pathways, verified on transcriptomics, proteomics and metabolomics level
● Extensive validation with multiple interventions (including the FDA approved drug)
● Metabolic overload-induced complications in multiple organs.
Time lines・Readout Paramters ; The Ldlr-/-.Leiden MASH mouse model
The Ldlr-/-.Leiden MASH model has been validated with many different interventions
| Intervention | Target | Publication |
|---|---|---|
| Caspase-1 inhibitor | Caspase 1 | Morrison, Int J Obes 2016 |
| Cenicriviroc | CCR2 & CCR5 | In preparation |
| Antioxidants (olive oil polyphenols) |
Oxidative stress | Luque-Sierra et al., Mol Nutr Food Res, 2018 Alvarez-Amor et al., Nature Sci Rep. 2021 |
| Rosi-, Pioglitazone | PPAR-γ | Mulder et al., Nat Sci Rep 2016 & in-house |
| Lanifibranor | PPARs | In preparation |
| EPA, DHA from krill oil | PPARs, oxylipins, resolvins | Gart et al., Nutrients 2021 |
| Obeticholic acid | FXR | Salic et al., PLoS One 2019 |
| Volixibat | IBAT | Mulder et al., Nat Sci Rep 2016 & in-house |
| Butyric acid | i.a. TGF-β fibrosis sign. | Arnoldussen et al., Int J Obes 2017 Gart et al., Biomedicines 2021 |
| Propionic acid | TCA cycle | Gart et al., FASEB J 2020 |
| Semaglutide | GLP-1 | Inia et al., Int J of Mol Sci 2023 |
| DGAT2 inhibitor | DAGT2 | In preparation |
| L-carnitine and nicotinamide riboside | β-oxidation, mitochondria | Salic and Gart et al., Int J Mol Sc 2019 |
| Casein hydrolysate | i.a. lipid modulators | Schoemaker et al., PLoS One 2017 |
| Branched chain amino acids | Mitochondria, TCA cycle | Gart et al., FASEB J. 2022 van den Hoek et al., Metabolism 2021 |
| Milk-fat globule membrane | Metabolism | Arnoldussen et al., Int J Obes 2021 |
| Natural cholesterol lowering compounds | Neutrophils | Morrison et al., Front Endocrinol 2021 |
| Resmetirom | THR-β | In preperation |
| Translation to Human | Translational Value | Head to head with 1099 patients |
●Download; latest publication list (pdf)
●Recent poster at AASLD 2022: The bispecific anti FGFR1/KLB agonist antibody bFKB1 attenuates non alcoholic steatohepatitis and atherosclerosis in Ldlr -/-.Leiden mice.
●Poster-1 at AASLD 2023: Temporal dynamics of metabolic dysfunctions in liver, adipose tissue and the gut during diet-induced NASH development in Ldlr-/-.Leiden mice.
●Poster-2 at AASLD 2023: Clinical translatability of Ldlr-/-.Leiden mouse model for NASH
Liver-on-a-Chip・ Cell model for liver fibrosis
We have developed a liver-on-a-chip model that exhibits MASH / liver fibrosis. By adding oleate and palmitate and later fructose, steatosis and steatohepatitis are induced. Adding also LPS leads to the induction of fibrosis.
This 3D liver spheroid model uses human hepatocytes, stellate cells and Kupffer cells. The model is sensitive to reference compounds such as pioglitazone and fenofibrate.
Summary:
・A diet-induced disease-mimicking 3D in vitro model, closely resembling the pathophysiology of liver steatosis and early fibrosis was developed;
・Liver spheroids composed of primary hepatocytes, stellate and Kupffer cells exhibit expected cell function for at least 21 days;
・The steatosis and steatohepatitis induced by fatty acids and fructose can be modulated by model drugs;
● Development of a disease-mimicking model for NASH and fibrosis in a triple cell-type, spheroid-based liver-on-chip platform with microfluidics.(Poster)
Biomarkers for MASLD・MASH
Fibrosis is the main determinant for mortality and is therefore an important clinical readout. Detection of fibrosis via a liver biopsy is still the golden standard but this is an invasive procedure with risk for complications, sampling and reading errors.
Fibrosis is a dynamic process and currently no validated diagnostic/prognostic methods exist, especially biomarkers which can detect the early onset and progression of fibrosis.
We aim to identify a mechanism-based biomarker panel which is prognostic for MASLD-related fibrosis and enables early identification of people at risk with active fibrogenesis.
By integration of mechanistic mouse studies and human clinical samples we identified a set of circulating proteins related to the active fibrogenesis process.
● Hepatic molecular signature in a translational NASH model as an early screening tool for novel NASH therapeutics.(Poster)
●(New) Early prediction of progressive fibrogenesis in NAFLD-NASH patients using blood-based biomarkers.(Poster)
● Optimized assays in Plasma, Serum, Breast milk, Fecal water, Saliva, etc validated in healthy subject or in patient groups.
