Hormone and Drug-Mediated Modulation of Glucose Metabolism in a Microscale Model of the Human Liver

Due to its central role in glucose homeostasis, the liver is an important target for drug development efforts for type 2 diabetes mellitus (T2DM). Significant differences across species in liver metabolism necessitate supplementation of animal data with assays designed to assess human-relevant responses. However, isolated primary human hepatocytes (PHHs) display a rapid decline in phenotypic functions in conventional monolayer formats. Cocultivation of PHHs with specific stromal cells, especially in micropatterned configurations, can stabilize some liver functions for ∼4 weeks in vitro. However, it remains unclear whether coculture approaches can stabilize glucose metabolism that can be modulated with hormones in PHHs. Thus, in this study, we compared commonly employed conventional culture formats and previously developed micropatterned cocultures (MPCCs) of cryopreserved PHHs and stromal fibroblasts for mRNA expression of key glucose metabolism genes (i.e., phosphoenolpyruvate carboxykinase-1 [PCK1]) and...

[1]  R G Shulman,et al.  Quantitation of hepatic glycogenolysis and gluconeogenesis in fasting humans with 13C NMR. , 1991, Science.

[2]  B. Viollet,et al.  Revisiting the mechanisms of metformin action in the liver. , 2013, Annales d'endocrinologie.

[3]  Yuichi Sugiyama,et al.  Involvement of Organic Cation Transporter 1 in Hepatic and Intestinal Distribution of Metformin , 2002, Journal of Pharmacology and Experimental Therapeutics.

[4]  R. Sherwin.  Role of Liver in Glucose Homeostasis , 1980, Diabetes Care.

[5]  Peter V. Henstock,et al.  Cellular imaging predictions of clinical drug-induced liver injury. , 2008, Toxicological sciences : an official journal of the Society of Toxicology.

[6]  André Guillouzo,et al.  Evolving concepts in liver tissue modeling and implications for in vitro toxicology , 2008, Expert opinion on drug metabolism & toxicology.

[7]  Tony J Collins,et al.  ImageJ for microscopy. , 2007, BioTechniques.

[8]  H. Green,et al.  Formation of a keratinizing epithelium in culture by a cloned cell line derived from a teratoma , 1975, Cell.

[9]  Adam S. Hayward,et al.  Recent advances in 2D and 3D in vitro systems using primary hepatocytes, alternative hepatocyte sources and non-parenchymal liver cells and their use in investigating mechanisms of hepatotoxicity, cell signaling and ADME , 2013, Archives of Toxicology.

[10]  X. Chu,et al.  Comparison of Immortalized Fa2N-4 Cells and Human Hepatocytes as in Vitro Models for Cytochrome P450 Induction , 2008, Drug Metabolism and Disposition.

[11]  S. Haffner,et al.  Liver markers and development of the metabolic syndrome: the insulin resistance atherosclerosis study. , 2005, Diabetes.

[12]  O. Cummings,et al.  Design and validation of a histological scoring system for nonalcoholic fatty liver disease , 2005, Hepatology.

[13]  M. Melis,et al.  Hydrophobic characterization of intracellular lipids in situ by Nile Red red/yellow emission ratio. , 2008, Micron.

[14]  K. Petersen,et al.  The role of hepatic lipids in hepatic insulin resistance and type 2 diabetes , 2014, Nature.

[15]  Korkut Uygun,et al.  A novel 3D liver organoid system for elucidation of hepatic glucose metabolism , 2012, Biotechnology and bioengineering.

[16]  Z. Kmieć,et al.  Cooperation of Liver Cells in Health and Disease , 2001, Advances in Anatomy Embryology and Cell Biology.

[17]  Melvin E Andersen,et al.  Long‐Term Stability of Primary Rat Hepatocytes in Micropatterned Cocultures , 2013, Journal of biochemical and molecular toxicology.

[18]  G. Baffy.  Kupffer cells in non-alcoholic fatty liver disease: the emerging view. , 2009, Journal of hepatology.

[19]  L. Adams,et al.  Nonalcoholic fatty liver disease and diabetes mellitus: pathogenesis and treatment , 2011, Nature Reviews Endocrinology.

[20]  K. Jungermann,et al.  Zonation of parenchymal and nonparenchymal metabolism in liver. , 1996, Annual review of nutrition.

[21]  Frank Stahl,et al.  Comparison of primary human hepatocytes and hepatoma cell line Hepg2 with regard to their biotransformation properties. , 2003, Drug metabolism and disposition: the biological fate of chemicals.

[22]  S. Donkin,et al.  Regulation of hepatic glucose metabolism by leptin in pig and rat primary hepatocyte cultures. , 2004, American journal of physiology. Regulatory, integrative and comparative physiology.

[23]  J. Clore,et al.  Nonalcoholic steatohepatitis: association of insulin resistance and mitochondrial abnormalities. , 2001, Gastroenterology.

[24]  P Smith,et al.  Concordance of the toxicity of pharmaceuticals in humans and in animals. , 2000, Regulatory toxicology and pharmacology : RTP.

[25]  H. Green,et al.  Seria cultivation of strains of human epidemal keratinocytes: the formation keratinizin colonies from single cell is , 1975, Cell.

[26]  R. Coleman,et al.  Hepatic triacylglycerol accumulation and insulin resistance This work was supported by National Institutes of Health Grants DK56598, DK59935, and DK56350. Published, JLR Papers in Press, November 6, 2008. , 2009, Journal of Lipid Research.

[27]  K. Cusi.  Nonalcoholic fatty liver disease in type 2 diabetes mellitus , 2009, Current opinion in endocrinology, diabetes, and obesity.

[28]  Sangeeta N. Bhatia,et al.  Persistent hepatitis C virus infection in microscale primary human hepatocyte cultures , 2010, Proceedings of the National Academy of Sciences.

[29]  A. Guillouzo,et al.  Maintenance and reversibility of active albumin secretion by adult rat hepatocytes co-cultured with another liver epithelial cell type. , 1983, Experimental cell research.

[30]  S. Yamada,et al.  The effects of dexamethasone on metabolic activity of hepatocytes in primary monolayer culture , 1980, In Vitro.

[31]  B. Bilir,et al.  Species differences in hepatocyte induction of CYP1A1 and CYP1A2 by omeprazole , 1999, Human & experimental toxicology.

[32]  G. Shulman,et al.  Mechanisms for Insulin Resistance: Common Threads and Missing Links , 2012, Cell.

[33]  Yvonne Will,et al.  Use of micropatterned cocultures to detect compounds that cause drug-induced liver injury in humans. , 2013, Toxicological sciences : an official journal of the Society of Toxicology.

[34]  Sean Koppe.  Obesity and the liver: nonalcoholic fatty liver disease. , 2014, Translational research : the journal of laboratory and clinical medicine.

[35]  J. Girard,et al.  Contribution of de novo fatty acid synthesis to hepatic steatosis and insulin resistance: lessons from genetically engineered mice. , 2008, The Journal of clinical investigation.

[36]  M. Rewers,et al.  Insulin sensitivity in subjects with type 2 diabetes. Relationship to cardiovascular risk factors: the Insulin Resistance Atherosclerosis Study. , 1999, Diabetes care.

[37]  G. Shulman,et al.  Niclosamide ethanolamine improves blood glycemic control and reduces hepatic steatosis in mice , 2014, Nature Medicine.

[38]  Thomas D. Schmittgen,et al.  Analyzing real-time PCR data by the comparative CT method , 2008, Nature Protocols.

[39]  S. Prato,et al.  Management of type 2 diabetes: new and future developments in treatment , 2011, The Lancet.

[40]  B. Robinson,et al.  3‐Mercaptopicolinic acid, a preferential inhibitor of the cytosolic phosphoenolpyruvate carboxykinase , 1975, FEBS letters.

[41]  J. Dubé,et al.  Depletion of Liver Kupffer Cells Prevents the Development of Diet-Induced Hepatic Steatosis and Insulin Resistance , 2009, Diabetes.

[42]  M L Yarmush,et al.  Effect of cell–cell interactions in preservation of cellular phenotype: cocultivation of hepatocytes and nonparenchymal cells , 1999, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[43]  Anna-Karin Sohlenius-Sternbeck,et al.  Determination of the hepatocellularity number for human, dog, rabbit, rat and mouse livers from protein concentration measurements , 2006 .

[44]  Melvin E. Andersen,et al.  Organotypic liver culture models: Meeting current challenges in toxicity testing , 2012, Critical reviews in toxicology.

[45]  Chaodong Wu,et al.  Reduction of hepatic glucose production as a therapeutic target in the treatment of diabetes. , 2005, Current drug targets. Immune, endocrine and metabolic disorders.