title: 3D engineered In vitro hepatospheroids for studying drug toxicity and metabolism.
abstract: Drug toxicity is one of the reasons for late stage drug attrition, because of hepatotoxicity. Various in vitro liver models like primary human hepatocytes, immortalized human hepatic cell lines, liver slices and microsomes have been used; but limited by viability, hepatic gene expression and function. The 3D-engineered construct of hepatocyte-like-cells (HLCs) differentiated from stem cells, may provide a limitless source of hepatocytes with improved reproducibility. Towards this end, we used hepatospheroids (diameter=50-80μm) differentiated from human-umbilical-cord-mesenchymal stem cells (hUC-MSCs) on 3D scaffold GEVAC (Gelatin-vinyl-acetate-copolymer) as in vitro model for studying drug metabolism/toxicity. Our data demonstrated that hUC-MSCs-derived-hepatospheroids cultured on GEVAC expressed significantly higher drug-metabolizing enzymes (CYPs) both at mRNA and activity level compared to 2D culture, using HR-LC/MS. We further showed that hepatospheroids convert phenacetin (by CYP1A2) and testosterone (by CYP3A4) to their human-specific metabolites acetaminophen and 6β-hydroxytestosterone with a predictive clearance rate of 0.011ml/h/106cells and 0.021ml/h/106cells respectively, according to first-order kinetics. Hepatotoxicity was confirmed by exposing hepatospheroids to ethanol and acetaminophen; ROS generation, cell viability, cytoskeleton structure, elevation of liver function enzymes, i.e. AST and ALT, was analyzed. To the best of our knowledge, this is the first report to use hUC-MSCs-derived-hepatospheroids on GEVAC as in vitro model for drug metabolism/toxicity study; which can replace the conventional 2D-models used in drug development.
authors: Department of Biological Sciences, Sunandan Divatia School of Science, SVKM'S NMIMS (Deemed-to-be ) University, V. L Mehta road, Vile Parle (West), Mumbai 400056, Maharashtra, India.
authors: Swati
authors: S
authors: Chitrangi
authors: Division of Tissue Engineering and Regeneration Technologies, Biomedical Technology Wing, Shree Chitra Tirunal Institute for Medical Sciences and Technology, Thiruvananthapuram 695012, Kerala, India.
authors: Prabha
authors: P
authors: Nair
authors: Department of Biological Sciences, Sunandan Divatia School of Science, SVKM'S NMIMS (Deemed-to-be ) University, V. L Mehta road, Vile Parle (West), Mumbai 400056, Maharashtra, India. Electronic address: aparna.khanna@nmims.edu.
authors: Aparna
authors: A
authors: Khanna
mesh_terms: D018929:Cell Culture Techniques; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D064420:Drug-Related Side Effects and Adverse Reactions; D005780:Gelatin; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D004364:Pharmaceutical Preparations / Q000378:metabolism; D010615:Phenacetin / Q000494:pharmacology; D011108:Polymers; D017382:Reactive Oxygen Species / Q000378:metabolism; D018874:Spheroids, Cellular / Q000187:drug effects / Q000378:metabolism; D013739:Testosterone / Q000494:pharmacology; D014470:Umbilical Cord / Q000166:cytology*; D014753:Vinyl Compounds
chemical_list: D004364:Pharmaceutical Preparations; D011108:Polymers; D017382:Reactive Oxygen Species; D014753:Vinyl Compounds; D013739:Testosterone; D005780:Gelatin; D003577:Cytochrome P-450 Enzyme System; D010615:Phenacetin; C011566:vinyl acetate
keywords: Drug metabolism; Hepatospheroid; Hepatotoxicity testing; Mesenchymal stem cell; Pharmacokinetics; Polymer scaffold