title: Bioprinted 3D Primary Liver Tissues Allow Assessment of Organ-Level Response to Clinical Drug Induced Toxicity In Vitro.
abstract: Modeling clinically relevant tissue responses using cell models poses a significant challenge for drug development, in particular for drug induced liver injury (DILI). This is mainly because existing liver models lack longevity and tissue-level complexity which limits their utility in predictive toxicology. In this study, we established and characterized novel bioprinted human liver tissue mimetics comprised of patient-derived hepatocytes and non-parenchymal cells in a defined architecture. Scaffold-free assembly of different cell types in an in vivo-relevant architecture allowed for histologic analysis that revealed distinct intercellular hepatocyte junctions, CD31+ endothelial networks, and desmin positive, smooth muscle actin negative quiescent stellates. Unlike what was seen in 2D hepatocyte cultures, the tissues maintained levels of ATP, Albumin as well as expression and drug-induced enzyme activity of Cytochrome P450s over 4 weeks in culture. To assess the ability of the 3D liver cultures to model tissue-level DILI, dose responses of Trovafloxacin, a drug whose hepatotoxic potential could not be assessed by standard pre-clinical models, were compared to the structurally related non-toxic drug Levofloxacin. Trovafloxacin induced significant, dose-dependent toxicity at clinically relevant doses (≤ 4uM). Interestingly, Trovafloxacin toxicity was observed without lipopolysaccharide stimulation and in the absence of resident macrophages in contrast to earlier reports. Together, these results demonstrate that 3D bioprinted liver tissues can both effectively model DILI and distinguish between highly related compounds with differential profile. Thus, the combination of patient-derived primary cells with bioprinting technology here for the first time demonstrates superior performance in terms of mimicking human drug response in a known target organ at the tissue level.
authors.affiliation: Organovo Holdings Inc., San Diego, CA, United States of America.
authors.forename: Deborah G
authors.identifier: http://orcid.org/0000-0002-1694-0617
authors.initials: DG
authors.lastname: Nguyen
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center, Basel, Switzerland.
authors.forename: Juergen
authors.initials: J
authors.lastname: Funk
authors.affiliation: Organovo Holdings Inc., San Diego, CA, United States of America.
authors.forename: Justin B
authors.initials: JB
authors.lastname: Robbins
authors.affiliation: Organovo Holdings Inc., San Diego, CA, United States of America.
authors.forename: Candace
authors.initials: C
authors.lastname: Crogan-Grundy
authors.affiliation: Organovo Holdings Inc., San Diego, CA, United States of America.
authors.forename: Sharon C
authors.initials: SC
authors.lastname: Presnell
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center, Basel, Switzerland.
authors.forename: Thomas
authors.initials: T
authors.lastname: Singer
authors.affiliation: Roche Pharmaceutical Research and Early Development, Roche Innovation Center, Basel, Switzerland.
authors.forename: Adrian B
authors.initials: AB
authors.lastname: Roth
mesh_terms: D000418:Albumins / Q000378:metabolism; D062028:Bioprinting; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury / Q000175:diagnosis*; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism; D064420:Drug-Related Side Effects and Adverse Reactions; D024841:Fluoroquinolones / Q000008:administration & dosage; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional; D064704:Levofloxacin / Q000008:administration & dosage; D008070:Lipopolysaccharides / Q000378:metabolism; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D009287:Naphthyridines / Q000008:administration & dosage; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism
chemical_list: D000418:Albumins; D024841:Fluoroquinolones; D008070:Lipopolysaccharides; D009287:Naphthyridines; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D064704:Levofloxacin; C080163:trovafloxacin; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human