scholarly journals Human Liver Spheroids as a Model to Study Aetiology and Treatment of Hepatic Fibrosis

Cells ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 964 ◽  
Author(s):  
Tracey Hurrell ◽  
Vlasia Kastrinou-Lampou ◽  
Achilleas Fardellas ◽  
Delilah F. G. Hendriks ◽  
Åsa Nordling ◽  
...  
Keyword(s):  
Human Liver ◽  
Progressive Disease ◽  
Human Hepatocytes ◽  
Alcoholic Fatty Liver ◽  
Drug Candidates ◽  
Alk5 Inhibitor ◽  
Hepatocyte Markers ◽  
Parenchymal Cells ◽  
Oxidant Effect ◽  
Alcoholic Fatty Liver Disease

Non-alcoholic fatty liver disease affects approximately one billion adults worldwide. Non-alcoholic steatohepatitis (NASH) is a progressive disease and underlies the advancement to liver fibrosis, cirrhosis, and hepatocellular carcinoma, for which there are no FDA-approved drug therapies. We developed a hetero-cellular spheroid system comprised of primary human hepatocytes (PHH) co-cultured with crude fractions of primary human liver non-parenchymal cells (NPC) from several matched or non-matched donors, to identify phenotypes with utility in investigating NASH pathogenesis and drug screening. Co-culture spheroids displayed stable expression of hepatocyte markers (albumin, CYP3A4) with the integration of stellate (vimentin, PDGFRβ), endothelial (vWF, PECAM1), and CD68-positive cells. Several co-culture spheroids developed a fibrotic phenotype either spontaneously, primarily observed in PNPLA3 mutant donors, or after challenge with free fatty acids (FFA), as determined by COL1A1 and αSMA expression. This phenotype, as well as TGFβ1 expression, was attenuated with an ALK5 inhibitor. Furthermore, CYP2E1, which has a strong pro-oxidant effect, was induced by NPCs and FFA. This system was used to evaluate the effects of anti-NASH drug candidates, which inhibited fibrillary deposition following 7 days of exposure. In conclusion, we suggest that this system is suitable for the evaluation of NASH pathogenesis and screening of anti-NASH drug candidates.

scholarly journals Lipid droplets disrupt mechanosensing in human hepatocytes

2020 ◽  
Author(s):  
LiKang Chin ◽  
Neil D. Theise ◽  
Abigail E. Loneker ◽  
Paul A. Janmey ◽  
Rebecca G. Wells
Keyword(s):  
Nuclear Localization ◽  
Lipid Droplets ◽  
Significant Risk ◽  
Focal Adhesions ◽  
Significant Risk Factor ◽  
Human Hepatocytes ◽  
Matrix Stiffness ◽  
Alcoholic Fatty Liver ◽  
Mechanical Signal ◽  
Alcoholic Fatty Liver Disease

ABSTRACTHepatocellular carcinoma (HCC) is the fourth leading cause of cancer death in the world. Although most cases occur in stiff, cirrhotic livers, and stiffness is a significant risk factor, HCC can also arise in non-cirrhotic livers in the setting of non-alcoholic fatty liver disease (NAFLD). We hypothesized that lipid droplets in NAFLD might apply mechanical forces to the nucleus, functioning as mechanical stressors akin to stiffness. We investigated the effect of lipid droplets on cellular mechanosensing and found that primary human hepatocytes loaded with the fatty acids oleate and linoleate exhibited decreased stiffness-induced cell spreading and disrupted focal adhesions and stress fibers. The presence of large lipid droplets in hepatocytes resulted in increased nuclear localization of the mechano-sensor Yes-associated protein (YAP). In cirrhotic livers from patients with NAFLD, hepatocytes filled with large lipid droplets showed significantly higher nuclear localization of YAP as compared to cells with small lipid droplets. This work suggests that lipid droplets induce a mechanical signal that disrupts the ability of the hepatocyte to sense its underlying matrix stiffness, and that the presence of lipid droplets can induce intracellular mechanical stresses.GRAPHICAL ABSTRACT

scholarly journals AKR1D1 is a novel regulator of metabolic phenotype in human hepatocytes and is dysregulated in non-alcoholic fatty liver disease

Metabolism ◽  
2019 ◽  
Vol 99 ◽  
pp. 67-80 ◽  
Author(s):  
Nikolaos Nikolaou ◽  
Laura L. Gathercole ◽  
Lea Marchand ◽  
Sara Althari ◽  
Niall J. Dempster ◽  
...  
Keyword(s):  
Liver Disease ◽  
Fatty Liver ◽  
Fatty Liver Disease ◽  
Human Hepatocytes ◽  
Metabolic Phenotype ◽  
Alcoholic Fatty Liver ◽  
Alcoholic Fatty Liver Disease

scholarly journals Fat causes necrosis and inflammation in parenchymal cells in human steatotic liver

2021 ◽  
Author(s):  
Eddie Wisse ◽  
Filip Braet ◽  
Gerald J. Shami ◽  
Bartlomiej Zapotoczny ◽  
Celien Vreuls ◽  
...  
Keyword(s):  
Single Cell ◽  
Alcoholic Fatty Liver ◽  
Human Liver Tissue ◽  
Steatotic Liver ◽  
Mallory Bodies ◽  
Human Livers ◽  
Original Cell ◽  
Parenchymal Cells ◽  
Alcoholic Fatty Liver Disease ◽  
Peripheral Cytoplasm

AbstractAdapted fixation methods for electron microscopy allowed us to study liver cell fine structure in 217 biopsies of intact human livers over the course of 10 years. The following novel observations and concepts arose: single fat droplets in parenchymal cells can grow to a volume four times larger than the original cell, thereby extremely marginalizing the cytoplasm with all organelles. Necrosis of single parenchymal cells, still containing one huge fat droplet, suggests death by fat in a process of single-cell steatonecrosis. In a later stage of single-cell steatonecrosis, neutrophils and erythrocytes surround the single fat droplet, forming an inflammatory fat follicle indicating the apparent onset of inflammation. Also, fat droplets frequently incorporate masses of filamentous fragments and other material, most probably representing Mallory substance. No other structure or material was found that could possibly represent Mallory bodies. We regularly observe the extrusion of huge fat droplets, traversing the peripheral cytoplasm of parenchymal cells, the Disse space and the endothelium. These fat droplets fill the sinusoid as a sinusoidal lipid embolus. In conclusion, adapted methods of fixation applied to human liver tissue revealed that single, huge fat droplets cause necrosis and inflammation in single parenchymal cells. Fat droplets also collect Mallory substance and give rise to sinusoidal fat emboli. Therefore, degreasing of the liver seems to be an essential therapeutic first step in the self-repairing of non-alcoholic fatty liver disease. This might directly reduce single-cell steatotic necrosis and inflammation as elements in non-alcoholic steatohepatitis progression.

scholarly journals Human hepatocyte PNPLA3 148M exacerbates rapid non-alcoholic fatty liver disease development in chimeric mice

2021 ◽  
Author(s):  
Mohammad Kabbani ◽  
Eleftherios Michailidis ◽  
Sandra Steensels ◽  
Clifton Fulmer ◽  
Joseph Luna ◽  
...  
Keyword(s):  
Liver Disease ◽  
Fatty Liver ◽  
Fatty Liver Disease ◽  
Human Genetics ◽  
Human Hepatocytes ◽  
Chimeric Mice ◽  
Human Donor ◽  
Alcoholic Fatty Liver ◽  
Alcoholic Fatty Liver Disease ◽  
Global Health Problem

Abstract Advanced non-alcoholic fatty liver disease (NAFLD) is a rapidly emerging global health problem associated with pre-disposing genetic polymorphisms, most strikingly an isoleucine to methionine substitution in patatin-like phospholipase domain-containing protein 3 (PNPLA3-I148M). Here, we study how human hepatocytes with PNPLA3 148I and 148M variants engrafted in the livers of chimeric mice respond to hypercaloric diets. As early as 4 weeks, mice developed dyslipidemia, impaired glucose tolerance, and steatohepatitis selectively in the human graft, followed by pericellular fibrosis after 8 weeks of hypercaloric feeding. The PNPLA3 148M variant, either from a homozygous 148M human donor or overexpressed in a homozygous 148I donor background, caused microvesicular and more severe steatosis. In these livers hepatocytes displayed frequent ballooning degeneration, resulting in more active steatohepatitis than in 148I livers. We conclude that PNPLA3 148M in human hepatocytes exacerbates NAFLD. These models will facilitate mechanistic studies into human genetics associated with advanced fatty liver diseases.

scholarly journals Combination of CCl4 with alcoholic and metabolic injuries mimics human liver fibrosis

2019 ◽  
Vol 317 (2) ◽  
pp. G182-G194 ◽  
Author(s):  
Maximilian Joseph Brol ◽  
Felicitas Rösch ◽  
Robert Schierwagen ◽  
Fernando Magdaleno ◽  
Frank Erhard Uschner ◽  
...  
Keyword(s):  
Liver Disease ◽  
Carbon Tetrachloride ◽  
Liver Fibrosis ◽  
Fatty Liver ◽  
Fatty Liver Disease ◽  
Human Liver ◽  
Alcoholic Fatty Liver ◽  
Significant Fibrosis ◽  
Alcoholic Fatty Liver Disease ◽  
Significant Liver Fibrosis

Metabolic and alcoholic liver injuries result in nonalcoholic (NAFLD) or alcoholic (ALD) fatty liver disease, respectively. In particular, presence of fibrosis in NAFLD and ALD requires treatment, but development of drugs is hampered by the lack of suitable models with significant fibrosis. The carbon tetrachloride (CCl4) liver fibrosis model does not reflect human NAFLD or ALD, but CCl4 may serve as a fibrosis accelerator in addition to another injury. Ethanol in drinking water (16%) or Western diet (WD) were administered for 7 wk in mice either alone or in combination with CCl4 intoxications. Extent of fibrosis, steatosis, and inflammation was assessed by histology, transcription, and biochemistry. Furthermore, transcription of fibrosis, proliferation, and inflammation-related genes was studied on human liver samples with fibrosis resulting from hepatitis C virus infection ( n = 7), NAFLD ( n = 8), or ALD ( n = 7). WD or ethanol alone induced only mild steatosis and inflammation. Combination of CCl4 and WD induced the most severe steatosis together with significant liver fibrosis and moderate inflammation. Combination of CCl4 and ethanol induced the strongest inflammation, with significant liver fibrosis and moderate steatosis. The relationship pattern between fibrosis, proliferation, and inflammation of human ALD was mostly similar in mice treated with CCl4 and ethanol. The combination of CCl4 intoxication with WD validates previous data suggesting it as an appropriate model for human nonalcoholic steatohepatitis. Especially, CCl4 plus ethanol for 7 wk induces ALD in mice, providing a model suitable for further basic research and drug testing. NEW & NOTEWORTHY Alcoholic fatty liver disease with significant fibrosis is generated within 7 wk using carbon tetrachloride as a fibrosis accelerator and administering gradually ethanol (up to 16%) in mice. The similarity in the pattern of steatosis, inflammation, and fibrosis involved in alcoholic fatty liver disease to those of the human condition renders this mouse model suitable as a preclinical model for drug development.

Non-alcoholic fatty liver disease: a metabolic burden promoting atherosclerosis

2020 ◽  
Vol 134 (13) ◽  
pp. 1775-1799 ◽  
Author(s):  
Lei Zhang ◽  
Zhi-Gang She ◽  
Hongliang Li ◽  
Xiao-Jing Zhang
Keyword(s):  
Liver Disease ◽  
Fatty Liver ◽  
Fatty Liver Disease ◽  
Cell Activation ◽  
Cell Damage ◽  
Alcoholic Fatty Liver ◽  
Drug Candidates ◽  
Metabolic Burden ◽  
Molecular Events ◽  
Alcoholic Fatty Liver Disease

Abstract Non-alcoholic fatty liver disease (NAFLD) has become the fastest growing chronic liver disease, with a prevalence of up to 25% worldwide. Individuals with NAFLD have a high risk of disease progression to cirrhosis, hepatocellular carcinoma (HCC), and liver failure. With the exception of intrahepatic burden, cardiovascular disease (CVD) and especially atherosclerosis (AS) are common complications of NAFLD. Furthermore, CVD is a major cause of death in NAFLD patients. Additionally, AS is a metabolic disorder highly associated with NAFLD, and individual NAFLD pathologies can greatly increase the risk of AS. It is increasingly clear that AS-associated endothelial cell damage, inflammatory cell activation, and smooth muscle cell proliferation are extensively impacted by NAFLD-induced systematic dyslipidemia, inflammation, oxidative stress, the production of hepatokines, and coagulations. In clinical trials, drug candidates for NAFLD management have displayed promising effects for the treatment of AS. In this review, we summarize the key molecular events and cellular factors contributing to the metabolic burden induced by NAFLD on AS, and discuss therapeutic strategies for the improvement of AS in individuals with NAFLD.

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