A Predictive 3D Multi-Scale Model of Biliary Fluid Dynamics in the Liver Lobule.

Bile, the central metabolic product of the liver, is transported by the bile canaliculi network. The impairment of bile flow in cholestatic liver diseases has urged a demand for insights into its regulation. Here, we developed a predictive 3D multi-scale model that simulates fluid dynamic properties successively from the subcellular to the tissue level. The model integrates the structure of the bile canalicular network in the mouse liver lobule, as determined by high-resolution confocal and serial block-face scanning electron microscopy, with measurements of bile transport by intravital microscopy. The combined experiment-theory approach revealed spatial heterogeneities of biliary geometry and hepatocyte transport activity. Based on this, our model predicts gradients of bile velocity and pressure in the liver lobule. Validation of the model predictions by pharmacological inhibition of Rho kinase demonstrated a requirement of canaliculi contractility for bile flow in vivo. Our model can be applied to functionally characterize liver diseases and quantitatively estimate biliary transport upon drug-induced liver injury.

[1]  Walter de Back,et al.  Morpheus: a user-friendly modeling environment for multiscale and multicellular systems biology , 2014, Bioinform..

[2]  N. Ballatori,et al.  Relation between biliary glutathione excretion and bile acid-independent bile flow. , 1989, The American journal of physiology.

[3]  L. Wong,et al.  Effect of aspirin on a subtoxic dose of 14C-acetaminophen in mice. , 1977, Journal of pharmaceutical sciences.

[4]  E. Heathcote.  Diagnosis and management of cholestatic liver disease. , 2007, Clinical gastroenterology and hepatology : the official clinical practice journal of the American Gastroenterological Association.

[5]  W. Lo,et al.  Apical membrane rupture and backward bile flooding in acetaminophen-induced hepatocyte necrosis , 2011, Cell Death and Disease.

[6]  W. J. Racz,et al.  Scanning electron microscopic examination of acetaminophen-induced hepatotoxicity and congestion in mice. , 1983, The American journal of pathology.

[7]  J. Boyer,et al.  Drug‐induced cholestasis , 2011, Hepatology.

[8]  Robert P Tonge,et al.  Genomics and proteomics analysis of acetaminophen toxicity in mouse liver. , 2002, Toxicological sciences : an official journal of the Society of Toxicology.

[9]  M H Zwietering,et al.  Characterization of uptake and hydrolysis of fluorescein diacetate and carboxyfluorescein diacetate by intracellular esterases in Saccharomyces cerevisiae, which result in accumulation of fluorescent product , 1995, Applied and environmental microbiology.

[10]  J. Boyer,et al.  Bile formation and secretion. , 2013, Comprehensive Physiology.

[11]  R. Tsien,et al.  Enhancing Serial Block-Face Scanning Electron Microscopy to Enable High Resolution 3-D Nanohistology of Cells and Tissues , 2010 .

[12]  Frank Bradke,et al.  Lifeact mice for studying F-actin dynamics , 2010, Nature Methods.

[13]  A. Guillouzo,et al.  Rho-kinase/myosin light chain kinase pathway plays a key role in the impairment of bile canaliculi dynamics induced by cholestatic drugs , 2016, Scientific Reports.

[14]  R. Lutz,et al.  Manometric changes during retrograde biliary infusion in mice. , 2000, American journal of physiology. Gastrointestinal and liver physiology.

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

[16]  J. Boyer,et al.  Canalicular bile secretion in man. Studies utilizing the biliary clearance of (14C)mannitol. , 1974, The Journal of clinical investigation.

[17]  Y. Kalaidzidis,et al.  A versatile pipeline for the multi-scale digital reconstruction and quantitative analysis of 3D tissue architecture , 2015, eLife.

[18]  Xiaoyu Luo,et al.  On the mechanical behavior of the human biliary system. , 2007, World journal of gastroenterology.

[19]  M. Fallon,et al.  Ca2+ waves are organized among hepatocytes in the intact organ. , 1995, The American journal of physiology.

[20]  H. Elias,et al.  A re-examination of the structure of the mammalian liver; the hepatic lobule and its relation to the vascular and biliary systems. , 1949, The American journal of anatomy.

[21]  K. Jungermann.  Metabolic Zonation of Liver Parenchyma , 1988, Seminars in liver disease.

[22]  T. Layden,et al.  Influence of bile acids on bile canalicular membrane morphology and the lobular gradient in canalicular size. , 1978, Laboratory investigation; a journal of technical methods and pathology.

[23]  C. Oshio,et al.  Contractility of bile canaliculi: implications for liver function. , 1981, Science.

[24]  Kenneth W. Dunn,et al.  Quantitative intravital microscopy of hepatic transport , 2012 .

[25]  C. Barth,et al.  The extracorporeal bile duct: a new model for determination of bile flow and bile composition in the intact rat. , 1978, Journal of lipid research.

[26]  V. Vollrath,et al.  Induction of the multispecific organic anion transporter (cMoat/mrp2) gene and biliary glutathione secretion by the herbicide 2,4,5-trichlorophenoxyacetic acid in the mouse liver. , 1999, The Biochemical journal.

[27]  Takeshi Imai,et al.  SeeDB: a simple and morphology-preserving optical clearing agent for neuronal circuit reconstruction , 2013, Nature Neuroscience.

[28]  Y. Sugiyama,et al.  Characterization of bile acid transport mediated by multidrug resistance associated protein 2 and bile salt export pump. , 2001, Biochimica et biophysica acta.

[29]  D. Meijer,et al.  Autoradiographic and kinetic demonstration of acinar heterogeneity of taurocholate transport. , 1982, The American journal of physiology.

[30]  J. Boyer,et al.  Molecular pathogenesis of cholestasis. , 2012, The New England journal of medicine.

[31]  Sarah Seifert,et al.  Rab5 is necessary for the biogenesis of the endolysosomal system in vivo , 2012, Nature.

[32]  Nathanson Mh,et al.  Calcium signaling mechanisms in liver in health and disease. , 1996 .

[33]  A. Miyazaki,et al.  Bile canalicular contraction in the isolated hepatocyte doublet is related to an increase in cytosolic free calcium ion concentration. , 2008, Liver.

[34]  D. Billington,et al.  Membranes and bile formation. Composition of several mammalian biles and their membrane-damaging properties. , 1979, The Biochemical journal.

[35]  C. V. Van Itallie,et al.  Liver kinase B1 regulates hepatocellular tight junction distribution and function in vivo , 2016, Hepatology.

[36]  John Skilling,et al.  Data analysis : a Bayesian tutorial , 1996 .

[37]  N. Ballatori,et al.  Glutathione as a primary osmotic driving force in hepatic bile formation. , 1992, The American journal of physiology.

[38]  W. Hardison,et al.  Modulation of hepatic biotransformation and biliary excretion of bile acid by age and sinusoidal bile acid load. , 1987, The American journal of physiology.

[39]  G. Mulder,et al.  Formation of chemically reactive metabolites of phenacetin and acetaminophen. , 1981, Advances in experimental medicine and biology.

[40]  R. Mathias.  Epithelial water transport in a balanced gradient system. , 1985, Biophysical journal.

[41]  R. Evers,et al.  Canalicular multispecific organic anion transporter/multidrug resistance protein 2 mediates low-affinity transport of reduced glutathione , 1999 .

[42]  K. Brouwer,et al.  Pharmacokinetics of 5 (and 6)-Carboxy-2′,7′-Dichlorofluorescein and Its Diacetate Promoiety in the Liver , 2003, Journal of Pharmacology and Experimental Therapeutics.

[43]  J. Hinson,et al.  Mechanisms of acetaminophen oxidation to N-acetyl-P-benzoquinone imine by horseradish peroxidase and cytochrome P-450. , 1987, The Journal of biological chemistry.

[44]  E. Guibert,et al.  Different hepatocytes express the cholesterol 7α‐hydroxylase gene during its circadian modulation in vivo , 1995, Hepatology.

[45]  J. Boyer,et al.  Canalicular bile flow and bile secretory pressure. Evidence for a non-bile salt dependent fraction in the isolated perfused rat liver. , 1970, Gastroenterology.

[46]  F. Lammert,et al.  EASL Clinical Practice Guidelines: management of cholestatic liver diseases. , 2009, Journal of hepatology.

[47]  Chen-Yuan Dong,et al.  Visualization of hepatobiliary excretory function by intravital multiphoton microscopy. , 2007, Journal of biomedical optics.

[48]  P. Milkiewicz,et al.  Plasma elimination of cholyl-lysyl-fluorescein (CLF): a pilot study in patients with liver cirrhosis. , 2000, Liver.

[49]  F. Suchy,et al.  Intrahepatic cholestasis: Summary of an American Association for the Study of Liver Diseases single‐topic conference , 2005, Hepatology.

[50]  N. Watanabe,et al.  Motility of bile canaliculi in the living animal: implications for bile flow , 1991, The Journal of cell biology.

[51]  A. Bader,et al.  Hepatotoxicity and hepatic metabolism of available drugs: current problems and possible solutions in preclinical stages , 2010, Expert opinion on drug metabolism & toxicology.

[52]  D. Rickman,et al.  Apc tumor suppressor gene is the "zonation-keeper" of mouse liver. , 2006, Developmental cell.

[53]  W. Hardison,et al.  Greater taurodeoxycholate biotransformation during backward perfusion of rat liver. , 1986, The American journal of physiology.

[54]  N. Tsukada,et al.  Bile canalicular contraction is coincident with reorganization of pericanalicular filaments and co-localization of actin and myosin-II. , 1993, Journal of Histochemistry and Cytochemistry.

[55]  M. J. Phillips,et al.  Ca2+ causes active contraction of bile canaliculi: direct evidence from microinjection studies. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[56]  Robert L. McIntosh,et al.  A COMPREHENSIVE TISSUE PROPERTIES DATABASE PROVIDED FOR THE THERMAL ASSESSMENT OF A HUMAN AT REST , 2010 .

[57]  Johannes E. Schindelin,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[58]  P. Baier,et al.  Zonation of Hepatic Bile Salt Transporters , 2006, Digestive Diseases and Sciences.